ES2794653T3 - Diagnostic apparatus on paper substrate, and related methods and systems - Google Patents

Abstract

An apparatus comprising: a hydrophobic substrate (102) having a first end (104) and a second end (106) opposite the first end (104); a detection zone (116) on a first surface (112) of the substrate (102), the detection zone (116) defining an area for detecting an analyte in a sample, the detection zone (116) comprising: a first electrode (120) and a second electrode (122) disposed on the first surface (112) of the substrate (102); and a layer (138) of hydrophilic ink disposed on the first and second electrodes (120, 122) and an area between the first and second electrodes (120, 122); and a channel (140) comprising hydrophilic ink disposed on the first surface (112) of the substrate (102), the channel (140) having an entrance section (142) adjacent the first end (104) of the substrate (112), a central section (144) and an outlet section (146) in contact with the layer (138) of hydrophilic ink, which is arranged on the sensor (118), channel (140) that serves to transfer a fluid sample from the inlet section (142) to the hydrophilic ink layer (138).

Description

d e s c r ip tio n

Diagnostic apparatus on paper substrate, and related methods and systems

Disclosure field

The present invention relates generally to diagnostic apparatus, and more particularly to paper substrate diagnostic apparatus, and related methods and systems.

Background

Immunoassays, clinical chemistry assays, and / or other medical diagnostic tests are generally performed using automated diagnostic analyzers that assess the presence, absence, and / or concentration of a target analyte in a given sample. These automated diagnostic analyzers employ multiple carousels and multiple pipetting mechanisms to automatically aspirate fluid and dispense it to different areas of the analyzer to perform diagnostic testing procedures. However, due to the cost, size, and complexity of these automated diagnostic analyzers, the analyzers are typically found in laboratories, where large quantities of samples are sent to the laboratory for analysis. Therefore, the analysis of a sample with an automatic diagnostic analyzer requires a greater amount of time before receiving the results.

Point-Of-Care (POC) tests, on the other hand, involve the ^use of relatively smaller and simpler medical devices that can perform diagnostic tests at the bedside and / or in a ward. emergencies. Some devices used for POC testing use small microfluidic chips that include a microfluidic channel and a sensor. A fluid sample can be inserted into the microfluidic channel and transferred to the sensor. The device is then inserted into a reader that communicates with the sensor and determines the test results. Known POC devices, however, are generally expensive to manufacture and have poor assay performance.

Document JP 2007 256237 discloses a microchemical chip capable of analyzing a small amount of liquid sample, by forming a flow channel with a metal oxide film that is arranged on a hydrophobic substrate and becomes ultra-hydrophilic when light affects it. A channel pattern of the metal oxide film, which becomes ultra-hydrophilic when ultraviolet light falls on it, is formed on the surface of the substrate made of a hydrophobic material using an ink jet head ^{or the} like, and a section is arranged inlet for the liquid sample to be analyzed at one end of the standard, and a detection section for testing the liquid sample is arranged at the other end.

DUNGCHAI W. et al. "ANXALYTICAL CHEMISTRY". (20090715), ^vo I. 81. n.014, doi: 10.1021 / ac9007573, ISSN 0003-2700, pages 5821 - 5826, XP055210134 [Y] 1-10 * page 5822, column 1, paragraph 2 - page 5823, column 1, paragraph 2, Figures 1,2 disclose electrochemical detection for microfluidic devices on paper. Photolithography is used to manufacture microfluidic channels on filter paper, and screen printing technology is used to manufacture electrodes on microfluidic devices on paper. Screen-printed electrodes on paper using cyclic voltammetry are provided to demonstrate the basic electrochemical performance of the system. The devices are used for the determination of glucose, lactate, and uric acid in biological samples using enzyme oxidase reactions (glucose oxidase, lactate oxidase, and uricase, respectively).

Document AU 2014 100086 discloses a mobile computing device, configured as a low-cost detection system, comprising a microprocessor (202), one or more memory components (206) comprising a program and an accessible data store for the microprocessor, an image sensor (2I6) configured to acquire images and transfer digital representations of the images to the program and data store, and an audio interface (218) configured to generate electrical signals, operating under the control of the microprocessor . The audio interface is connectable, in ^use , to electrical input terminals of a chemical sensor (100) comprising a microfluidic layer (102) of paper loaded with active electrochemiluminescence (ECL, ElectroChemiLumescence) molecules ^or a co-reactant within a zone detection, and a flat circuit layer (104) comprising a working electrode (118) in contact with the detection zone of the paper microfluidic layer.

LIANA D. D. et al. SENSORS, voI. 12, no 12, pages 11505 - 11526, XP055236777, page 11507, paragraph 2 - page 11510, paragraph 4, page 11514, paragraph 1 - page 11516, paragraph 3, page 11517, paragraph 2 and figures 1, 2, 6, 7 disclose paper-based sensors for manufacturing simple, low-cost, portable and disposable analytical devices for application areas including clinical diagnostics, food quality control, and environmental monitoring. The authors mention the use of hydrophobic photoresist (FIG. 1) or wax printing (FIG. 2) to create a microfluidic channel in the paper. So when a liquid is introduced into the microfluidic channel, the liquid moves through the paper matrix by capillary flow action.

JENKINS G. et al. "MICROFLUIDICS AND NANOFLUIDICS", voI. 19, no 2. Pages 251 - 261, document XP055237188 describes a process for producing microfluidic channels on paper using agarose gel printing. In FIG. 6, a gelling agarose is shown which is printed on the paper and then cooled to form a gel. The paper is then treated and washed with hot water to melt and dissolve the agarose gel, thus leaving a hydrophilic channel in the paper.

Brief description of the drawings

FIG. 1 is a perspective view of an illustrative substrate diagnostic apparatus or device constructed in accordance with the teachings of the present disclosure.

FIGS. 2A-2F are cross-sectional views taken along line A-A in FIG. 1 during different stages of forming the illustrative substrate diagnostic device of FIG. 1.

FIG. 3A is a cross-sectional view taken along line B-B in FIG. 1 of the illustrative substrate diagnostic device of FIG. 1.

FIG. 3B is a cross-sectional view taken along line C-C in FIG. 1 of the illustrative substrate diagnostic device of FIG. 1.

FIG. 4 is an illustrative system that includes the illustrative substrate diagnostic device of FIG. 1 and an illustrative reader for use with the illustrative substrate diagnostic device.

FIG. 5 is a diagram of an illustrative assembly for constructing the illustrative substrate diagnostic device of FIG. 1.

FIG. 6 is a block diagram of an illustrative processing system for the illustrative assembly of FIG. 5.

FIG. 7 is a perspective view of an alternative illustrative substrate diagnostic device constructed, which is not in accordance with the present invention.

FIG. 8 is a cross-sectional view taken along line D-D in FIG. 7 of the illustrative substrate diagnostic device of FIG. 7.

FIG. 9 is a diagram of an illustrative assembly for creating the illustrative substrate diagnostic device of FIG. 7.

FIG. 10 is a block diagram of an illustrative processing system for the illustrative assembly of FIG. 9.

FIG. 11 is a perspective view of another alternative illustrative substrate diagnostic device constructed in accordance with the teachings of the present disclosure.

FIG. 12 is a cross-sectional view taken along line E-E in FIG. 11 of the illustrative substrate diagnostic device of FIG. eleven.

FIG. 13 is a flow chart of an illustrative method that can be used to implement the illustrative assemblies disclosed herein and construct the illustrative substrate diagnostic devices of FIGS. 1 and 11.

FIG. 14 is a flow chart of an alternative illustrative method that can be used to implement the illustrative assemblies disclosed herein and construct the illustrative substrate diagnostic device of FIG. 7.

FIG. 15 is an illustrative processor platform that can be used to implement the illustrative methods, systems, and / or apparatus disclosed herein.

Figures are not to scale. Instead, multiple clear layers and regions can expand the thickness of the layers in the drawings. Where possible, the same reference numbers will be used throughout the drawing (s) and the accompanying written description to refer to the same or similar parts.

Detailed description

T ^he immunoassays (IA, ImmunAssays), clinical chemistry assays (CC, Clinical Chemistry) and / or other medical diagnostic tests (eg, hematology) commonly done with automatic analyzers in a laboratory diagnosis. AI and QC tests are used to analyze a sample or biological sample such as, for example, blood, urine, plasma, etc. to determine the presence, absence, and / or concentration of an article of interest or target (eg, an analyte). The article of interest or analyte may include, for example, a specific region of DNA, mitochondrial DNA, a specific region of RNA, messenger RNA, transfer RNA, mitochondrial RNA, a fragment, a complement, a peptide, a polypeptide, a enzyme, a prion, a protein, an antibody, an antigen, an allergen, a part of a biological entity such as a cell ^or a virion, a surface protein, and / or functional equivalents of the foregoing. Samples such as body fluids from a patient (eg, serum, whole blood, urine, swabs, plasma, cerebrospinal fluid, lymphatic fluids, tissue solids) can be analyzed using a number of different tests to provide information about the patient's health. In general, the analysis of a test sample involves the reaction of the test sample with one ^or more reagents with respect to one or more analytes. The reaction mixtures are analyzed with an apparatus for one or more characteristics such as, for example, the presence, absence and / or concentration of a certain analyte in the test sample. For example, the apparatus disclosed below can be used to analyze analytes related to cancer, thyroid problems, fertility, HIV ^or I ^will heart problems.

Due to the cost, size, and complexity of automated diagnostic analyzers, analyzers they are typically used in laboratories to analyze large batches of samples (eg 400 samples). The samples are sent to the lab, the tests are done, and the results are sent back to the clinic ^or hospital. Therefore, testing through an automated diagnostic analyzer in a laboratory generally requires a longer period of time before the test results are received.

Point-of-care (POC) testing, on the other hand, involves the ^use of simple, relatively small devices that can be used at the bedside or in an emergency room for faster results (for example, within a few minutes). A home pregnancy test is an example of a POC test. AI and QC testing can also be done through small POC devices. In general, known POC diagnostic devices typically include a small chip or substrate that has a detection component for detecting a reaction between the sample and a reagent and a microfluidic component for the displacement of the sample and other liquids around the chip. L ^os devices typically include a reagent that interacts with the sample and a reader to determine test results. L ^os devices move the fluid sample through flow channels and to the sensor which can measure the reaction.

Some known devices have a plurality of channels that are machined or stamped into the surface of the chips. However, the manufacture of these complex, relatively small channels leads to higher cost manufacturing processes. Some detection devices use magnetic actuation, digital microfluidics (eg, electrowetting), or acoustic fluids to move the sample through the device. These techniques also result in a more expensive manufacturing process. As a result, known POC diagnostic devices are generally expensive to manufacture and are therefore expensive to use.

The present invention relates to consumable diagnostic devices that are manufactured using less expensive material, cheaper manufacturing processes, and fewer manufacturing steps resulting in a relatively less expensive diagnostic device. As a result, the disclosed substrate diagnostic devices are relatively easier and less expensive to manufacture.

L ^os diagnostic devices of the present invention include a microfluidic component and a detection component (eg a sensor). The diagnostic device of the present invention includes a substrate having a hydrophobic coating or hydrophobic properties that repel fluid. A sensor, including one or more electrodes, is disposed on a surface (eg, a top surface) of the paper substrate, and a material suspension layer is disposed on the top surface of the paper substrate, and on and around to define a detection zone. In this example, the suspension of material is hydrophilic and includes particles such as, for example, microbeads ^or nanobeads that form a porous structure that absorbs ^or captures fluids through a network of said particles. To transfer a fluid sample to the detection zone, the illustrative device also includes a material suspension channel that is disposed on the upper surface of the paper substrate and leads from one end of the paper substrate to the detection zone. When a fluid sample is deposited at the end of the channel, the sample is captured (eg, through capillary action) down the channel and into the top layer of the sensor. In some examples, a reagent is disposed on top of the sensor (eg, between a sensor electrode and the suspension layer of material). In this way, the sample can react with the reagent in the detection zone and the sensor can measure the electrical properties generated by it. In addition, a sealing layer ^or fluid impermeable layer is arranged over the channel and the detection zone to form a leak-free path between the upper surface of the paper substrate (eg, a lower boundary) and the sealing layer. (for example, an upper limit). The sealing layer reduces evaporation of the fluid sample and also reduces the risk of contamination.

The apparatus of the present invention is defined in claim 1. A method of manufacturing the apparatus of the present invention is defined in claim 11.

Illustrative systems and methods for constructing or fabricating the illustrative substrate diagnostic device are also disclosed herein. An illustrative system includes a reel-to-reel assembly (eg, coil-fed continuous process). In this example, a roll of substrate is unwound from a spool at one end of the set and wound at the other end of the set. In one ^or more printing steps, the components (eg, sensor, layer, channel, reagent, etc.) of the diagnostic device are printed on the top surface of the substrate. For example, a conductive ink can be printed on the substrate to form the sensor electrodes. In some examples, the electrodes are printed onto the substrate by inkjet printing, gravure printing, ^or flexographic printing. The layer and the suspension channel of the material (eg, hydrophilic ink) can be printed on the upper surface of the substrate by rotary screen printing. The sealing layer can also be printed on the layer and the channel by rotary screen printing. In this way, the microfluidic component and the detection component of the device can be printed on top of the hydrophobic substrate, instead of using complex lithography ^or machining processes such as those employed by known devices. As a result, the substrate diagnostic device is less expensive to manufacture and is therefore a more cost-effective disposable POC device.

An illustrative apparatus disclosed herein includes a hydrophobic substrate having a first end and a second end opposite the first end. The apparatus includes a detection zone on a first surface of the substrate, the detection zone defining an area for detecting an analyte in a sample, the detection zone comprising a first electrode and a second electrode disposed on the first surface of the substrate and a hydrophilic ink layer arranged on the two electrodes and an area between the first and second electrodes. The apparatus also includes a channel comprising hydrophilic ink disposed on the first surface of the substrate, the channel having an inlet section adjacent the first end of the substrate, a central section, and an outlet section in contact with the hydrophilic ink layer. The channel is for transferring a fluid sample from the inlet section to the hydrophilic ink layer.

In some examples, the apparatus includes a fluid impermeable layer embossed on the substrate to cover the detection zone, the central section of the channel, and the outlet section of the channel. In some examples, the fluid impermeable layer is in contact with the first surface of the substrate near the channel and the hydrophilic ink layer. In some examples, the fluid impermeable layer comprises nanofibrillar cellulose (NFC, Nanofiber Cellulose) in a water-based suspension. In some examples, the fluid-impermeable layer is a hydrophobic ink that must be deposited at least by means of flexographic printing, screen printing, stencil printing ^or inkjet printing.

In some examples, the apparatus includes a reagent layer disposed on an upper surface of the first electrode, between the first electrode and the hydrophilic ink layer. In some examples, the first electrode is functionalized with a reagent.

In some examples, the hydrophilic ink comprises microbeads ^or nanobeads and a binder to adhere the microbeads ^or nanobeads to the first surface of the substrate. In some of these examples, the binder comprises at least one of polyvinyl chloride (PVC), polyvinylpyrrolidone (PVP) ^or nanofibrillar cellulose (NFC). In some examples, the apparatus includes a first contact disposed on the upper surface of the substrate, the first contact being electrically coupled to the first electrode, a second contact disposed on the upper surface of the substrate, the second contact being electrically coupled to the second electrode and a reader. In some of these examples, the reader includes a slot to receive at least a portion of the substrate that includes the first contact and the second contact, electrical connectors to receive signals from the first contact and the second contact, and a processor to determine one or more between amperometric, voltammetric or potentiometric measurements to detect a biomolecular interaction that occurs in the detection zone through the first and second electrodes. In some examples, the sensor further comprises a third electrode, and the processor must detect the biomolecular interaction by comparing (1) a difference in the first and second electrodes; and (2) a difference in the second and third electrodes. In some examples, the detection zone comprises a plurality of additional electrodes, and the processor must determine a plurality of measurements using the plurality of additional electrodes.

In some examples, the substrate comprises paper. In some examples, the substrate is flexible and printable.

Herein, an illustrative method of manufacturing a substrate diagnostic device is disclosed. The illustrative method includes depositing a sensor on an upper surface of the substrate, the substrate having a first end and a second end opposite the first end, depositing a hydrophilic ink layer on the sensor, and depositing a hydrophilic ink path on the upper surface of the substratum. The pathway leads from an area adjacent to the first end of the substrate to the hydrophilic ink layer.

In some examples, the method includes depositing a hydrophobic ink layer on top of the lane and the hydrophilic ink layer. In some examples, the hydrophobic ink comprises nanofibrillar cellulose (NFC) in a water-based suspension. In some examples, the deposition of the hydrophobic ink layer includes printing the hydrophobic ink using at least one of a flexographic printer, a screen printer, a stencil printer, ^or an inkjet printer.

In some examples, the sensor includes a first electrode and a second electrode, and deposition of the sensor includes printing the first electrode and the second electrode on the upper surface of the substrate. In some of these examples, the method includes depositing a reagent layer on the first electrode. In some examples, the reagent layer is disposed between the first electrode and the hydrophilic ink layer. In some examples, the first electrode is functionalized with a reagent.

In some examples, the deposition of the hydrophilic ink layer and the deposition of the hydrophilic ink path include printing the hydrophilic ink using at least one of a flexographic printer, a screen printer, a stencil printer ^or a printer of inkjet.

Turning now to the figures, FIG. 1 illustrates an illustrative substrate-based diagnostic device 100 used to measure and / or detect the presence, absence and / or concentration of an article of interest ^or analyte in a fluid sample. The illustrative device 100 produces results in a relatively short period of time, and the device 100 can be manufactured relatively inexpensively. In some examples, an AI test or a QC test is performed on device 100, depending on the type of reagent used and the detection method used. To provide a supporting surface for the detection component (s) and microfluidic components (described in detail below), device 100 includes a substrate or a support sheet 102. In the illustrated example, substrate 102 comprises a strip ^or sheet of paper. Illustrative paper substrate 102 is hydrophobic (eg, fluid impervious, fluid resistant) and repels fluids ^or reduces the amount of fluid that is permeable to paper substrate 102. In some examples, paper substrate 102 It is made of hydrophobic material (for example, trees and / or chemicals with hydrophobic properties). In other examples, the paper substrate 102 has a porous structure and is treated with a hydrophobic coating that prevents fluids from penetrating the porous structure of the paper substrate 102.

In the illustrated example, the paper substrate 102 has a first end 104, a second end 106 opposite the first end 104, a first side 108, a second side 110 opposite the first side 108, a top surface 112, and a bottom surface 114 opposite top surface l l2. Upper surface 112 defines a support surface for the microfluidic component (s) and the sensing component (s). In the illustrated example, the paper substrate 102 is substantially rectangular. However, in other examples, the paper substrate 102 may have another shape such as, for example, a square, a triangle, a circle, an ellipse, an irregular shape, ^or any other shape capable of supporting the microfluidic components and / or detection systems disclosed herein.

In an illustrative operation, a fluid sample is pipetted or otherwise deposited at or near the first end 104 of the substrate at or near an opening 142 of a pathway or channel 140. In the illustrated example, channel 140 is arranged on the top surface 112 of the paper substrate 102. The illustrative channel 140 includes a first end 142 (eg, a sample deposit area, an inlet), an intermediate section 144, and a second end 146 (eg, an outlet ). In other examples, there may be more than one channel. Channel 140 is used to transfer the sample to a detection zone 116, which is disclosed in more detail below. To transfer or move a fluid sample to detection zone 116, illustrative channel 140 comprises suspended material, which may be in the form of a network of particles, a hydrophilic mesh, and / or a hydrophilic ink. Illustrative channel 140 hydrophilic ink captures the fluid sample by capillary action from one end of channel 140 to the other end of channel 140. Second end 146 is in contact with a layer 138 of suspended material (eg, hydrophilic mesh and / or hydrophilic ink) from the detection zone 116. As a result, a fluid passing through the channel 140 from the first end 142 of the channel 140 through the central section 144 and through the second end 146 moves into the layer 138 of the detection zone 116, where the sample comes into contact with one ^or more reagents and electrodes 120, 122, 124, as provided in greater detail below.

The hydrophilic mesh ^or ink layer 138 allows the sample to move through the substrate 102 in the detection zone 116 to contact a sensor 118, which includes the electrodes 120, 122, 124, during a reaction. Layer 138 is disposed over sensor 118 and the area on top surface 112 surrounding sensor 118. In the illustrated example, layer 138 covers electrodes 120, 122, 124 and the area surrounding electrodes 120, 122, 124. In some examples, layer 138 may only cover a portion of electrodes 120, 122, 124.

The hydrophilic ink ^or material suspension disposed in channel 140 and layer 138 includes a suspension of particles such as, for example, microbeads (e.g. 25-36 micron (pm) silica beads, 10pm for aqueous solutions) or nanobeads, which create a porous and immobile structure (for example, a network or matrix of hydrophilic particles). Some of the particles can be hydrophilic and others can be hydrophobic, and the particles can vary in dimension and size. In some examples, the particles in the suspension of material are greater than 20 µm and less than 100 µm in diameter. In other examples, particles of another size diameter are used. In some examples, the particles are water-based for biological compatibility. The resulting hydrophilic structure absorbs fluids (eg, by uptake or capillary action). As a result, layer 138 absorbs the fluid sample so that the sample contacts (eg, moistens) electrodes 120, 122, 124 of sensor 118.

In some examples, the hydrophilic ink also includes binders that mix with the particles to aid bonding (eg, by adhesion) of the hydrophilic ink to the 102 paper substrate. In some examples, the binders include polyvinyl chloride (PVC ), polyvinylpyrrolidone (PVP) and / or nanofibrillar cellulose (NFC). In other examples, other types of binders can be used. L ^os binders and particles can be mixed in varying proportions , and in some examples, can be mixed with additional solvents. The proportions or percentages of binders and particles affect the properties (eg, pick-up rate) of the hydrophilic ink. Thus, the composition of the hydrophilic ink ^or material suspension in channel 140 can be varied to control fluid flow. Other factors that can also be used to control fluid flow include the characteristics of the particles, including, for example, the hydrophilicity and / or dimensions of the particles, the type of binder, the composition percentage of the suspension of material. (the ratio of particles to the volume of the fluid) and / or the dimensions of the channel, including width, length and / or height.

In some examples, the hydrophilic ink in channel 140 is the type of hydrophilic ink used in layer 138. In others For example, the hydrophilic ink in channel 140 is different from the hydrophilic ink in layer 138. For example, the hydrophilic ink in channel 140 may include a different ratio of particles and binders than the hydrophilic ink in layer 138 (for example, giving place at different uptake speeds.

In the illustrated example, the hydrophilic ink from channel 140 and / or layer 138 is printed (eg, stamped) on device 100 using a printing process such as, for example, gravure printing, flexographic printing, screen printing, rotary screen printing and / or ink jet printing. In other examples, other suitable types of printing processes may be used to print the hydrophilic ink on the paper substrate 102 to form channel 140 and / or layer 138. In some examples, channel 140 and layer 138 they are printed on the paper substrate 102 during the same printing process. In some examples, the type of printing process is determined based on the desired dimension (eg, thickness, width, and / or length) of layer 138. For example, in some examples, flexographic printing it allows a relatively finer resolution of the canal structures and is compatible with particles smaller than 10 pm. In some examples, screen printing or stencil printing produces larger channel structures and is compatible with a relatively larger range of particle sizes (eg depending on the characteristics of the printing mesh used).

In the illustrated example, device 100 includes detection zone 116 disposed on top surface 112 of sheet of paper 102. Detection zone 116 defines a location where a reaction occurs (for example, for the fluid sample scanned by an electronic reader). Detection zone 116 includes sensor 118 for detecting a reaction between the sample and one or more reagent / s. The detected information is used to determine, for example, the presence, absence and / or concentration of the target analyte in the sample. In the illustrated example, sensor 118 is an electrochemical sensor and includes a first electrode 120 (eg, a working electrode) and a second electrode 122 (eg, a counter electrode, a ground electrode) that are disposed on the surface. top 112 of the paper substrate 102. Biomolecular interactions that occur on the surface of the electrodes can be measured using electrical and / or electrochemical methods, including amperometric, voltammetric and / or potentiometric techniques. T ^he results electrical and / or electrochemical occurring at the two electrodes 120, 122 may be correlated with the presence, absence and / or concentration of the target analyte in the sample. In the illustrated example, a third electrode 124 (eg, a reference electrode) is also disposed on the upper surface 112 of the paper substrate 102. In some examples, the first electrode 120 is a working electrode, the second electrode 122 it is a ground or counter electrode, and the third electrode 124 is a reference electrode. When measuring the difference between the first and second electrodes 120, 122 (for example, the working electrode / counter electrode pair) and the second and third electrodes 122, 124 (for example, the reference / counter electrode pair), A measurement can be made to eliminate noise and / or other interference. In some examples, only two electrodes are implemented such as, for example, a working electrode and a counter electrode (for example, ground). In still other examples, multiple electrodes are used (eg, four, six, nine, etc.). In such an example, multiple measurements can be performed on the same device to detect multiple analytes.

In the illustrated example, device 100 employs electrical-type sensing via sensor 118. However, in other examples, other types of sensors may be implemented such as, for example, an optical sensor, a magnetic sensor, or a mechanical sensor. .

In some examples, the one ^or more of the electrodes 120, 122, 124 comprise gold (A ^u ), gold on silver chloride (AgCl), carbon (C) and / or silver / silver chloride (Ag / AgCl). In some examples, one or more of the electrodes 120, 122, 124 are printed on the upper surface 112 of the paper substrate 102 using conductive ink. For example, one ^or more of the electrodes 120, 122, 124 can be printed on the device using gravure printing, flexographic printing, screen printing, rotary screen printing and / or ink jet printing. In other examples, other suitable types of printing processes may be employed to print the conductive ink on the paper substrate 102.

To enable an electronic reader (disclosed in detail below) to electrically communicate with electrodes 120, 122, 124, device 100 includes one or more contacts. In the illustrated example, a first trace 126 (eg, a wire, a cable, etc.) and a first contact 128 are disposed on the upper surface 112 of the paper substrate 102. The first trace 126 electrically couples the first electrode 120 with the first contact 128. In the illustrated example, the first contact 128 is located near the second end 106 of the sheet of paper 102. However, in other examples, the first contact 128 may be disposed at other locations on the sheet of paper. paper 102.

In the illustrated example, a second trace 130 and a second contact 132 are arranged on the upper surface 112 of the sheet of paper 102 for the second electrode 122, and a third trace 134 and a third contact 136 are arranged on the upper surface 112 of the paper sheet 102 for the third electrode 124. In the illustrated example, the three contacts 128, 132, 136 are arranged in a row near the second end 106 of the paper sheet 102. As a result, the second end 106 of the Device can be inserted into a reader that has pins that connect to contacts 128, 132, 136. In some examples, traces 126, 130, 134 and contacts 128, 132, 136 are printed on top surface 112 of the substrate of paper 102 using printing processes similar to those for electrodes 120, 122, 124 disclosed above.

When performing AI and QC tests, one ^or more reagents are commonly used to interact with the sample. In some examples, one or more reagents (Illustrated in FIG. 2D) are disposed on top of the first electrode 120 (eg, the working electrode). As the sample comes into contact with the first electrode 120, the sample interacts with the reagent and the biomolecular interaction causes a change in volts, amps and / or resistance that can be measured across the first and second electrodes 120, 122 For example, sensor 118 may be a nanowire Field Effect Transistor (FET) sensor. In such an example, the FET inlet can be functionalized with the reagent and the electrochemical reaction between the sample at the inlet can be measured. The resulting change in electrical activity can be correlated and / or used to determine the presence, absence, and / or concentration of the target analyte in the sample (eg, by comparing the results with the reference data). In other examples, one ^or more reagents are functionalized directly on the material of the first electrode 120. In other examples, one or more reagents can be dispensed (eg, by printing) onto the top surface 112 of the paper substrate 112 (eg , below channel 140 ^or layer 138). As it is captured by channel 140 and / or layer 138, the sample reacts with one ^or more reagents.

In some examples, analyte detection can be performed using tagless detection such as, for example, impedance, capacitance, or resistance techniques that recognize the analyte of interest. Additionally or alternatively, a second binding reagent that recognizes a different region of the analyte of interest can be employed. In some examples, the second binding reagent improves detection specificity, enhances detection sensitivity, and / or improves a dynamic range of the assay. The second binding reagent can be conjugated to a label that includes conductive particles (for example, metallic colloids such as gold, silver, copper, platinum, carbon microparticles and / or carbon nanoparticles and / or conductive polymer particles) , quantum dots, latex particles, polymeric particles, magnetic particles (for example, magnetic microparticles and / or magnetic nanoparticles). In some examples, detection is based on the ^use of redox mediators and an enzyme catalyst that is deposited on the surface of the electrode (eg, the surface of the first electrode 120).

In some examples, to protect exposed conductive traces, a dielectric barrier material may be deposited (eg, by printing) in a specified pattern. In some examples, a dielectric ink is used, and the dielectric ink anchors and seals the hydrophilic ink from channel 140 and / or the hydrophilic ink from layer 138 to substrate 102. In some examples, the dielectric ink defines the exposed reaction volumes to sensor 118. In other examples, a waterproof membrane ^or resist layer can be placed in the desired location and sealed by lamination.

To prevent contamination and / or evaporation of the fluid sample as it moves through channel 140 and layer 138 and interacts with reagents in detection zone 116, illustrative device 100 includes sealing layer 148, which is disposed over channel 140 and layer 138. Seal layer 148 creates a fluid impervious ( ^or hydrophobic) surface on top of the hydrophilic ink in channel 140 and layer 138. In the illustrated example, the layer Seal 148 is not disposed above first end 142 of channel 140, so fluid sample can be deposited at first end 142 of channel 140 over the hydrophilic ink of channel 140. In other examples, the entire channel 140 is covered with seal layer 148 and an opening ^or hole is defined in seal layer 148 to allow the fluid sample to contact the hydrophilic ink in channel 140.

In the illustrated example, the sealing layer 148 is in contact with the upper surface 112 of the paper substrate 102 on the immediate sides of the channel 140 and the layer 138 of the detection zone 116. As a result, the sealing layer 148 forms or defines a sealed (eg, leak-free) pathway between the seal layer 148 and the top surface 112 of the paper substrate 102. In some examples, the seal layer 148 is an ink that is printed on the channel 140 and detection zone 116. In some examples, the ink includes NFC in a water-based suspension. In some examples, the ink seal layer 148 is printed on the device by flexographic printing, screen printing, stencil printing, and / or inkjet printing. In other examples, other types of printing processes can be used. In the illustrated example, seal layer 148 is not disposed over the remainder of the paper substrate 102 away from channel 140 and layer 138. As a result, less material is used to plug ^or seal channel 14 and layer 138. However, in other examples, the seal layer may be printed over the entire 102 paper substrate.

In some examples, the composition of the conductive ink / s, the sealing layer (e.g., hydrophobic ink / s, hydrophilic ink / s, and / or dielectric ink / s) may depend of the properties of the substrate 102 and / or of the printing processes used. For example, the conductive ink may need to be cured or dried after deposition (eg, via ultraviolet light, heat, infrared (IR), ^or air drying) depending on the ink composition and / or substrate tolerances.

FIGS. 2A-2F are cross-sectional views of device 100 taken along line AA in FIG 1. FIGS. 2A-2F illustrate illustrative steps for fabricating ^or constructing device 100. It should be noted that the figures are not necessarily to scale. In fact, elements in the figures have been exaggerated for illustrative purposes. Illustrative paper substrate 102 is illustrated in FIG. 2A. In some examples, the paper substrate 102 is pre-processed as a hydrophobic paper sheet. For example, the paper substrate 102 can be made from materials and / or chemicals that give rise to a hydrophobic paper substrate. In other examples, the paper substrate 102 may be a more traditional piece of porous paper that is treated with a hydrophobic coating ^or layer.

In the illustrated example of FIG. 2B, the first and second electrodes 120, 122 of the sensor 118 (FIG. 1) are disposed on the upper surface 112 of the paper substrate 102. In some examples, the first and second electrodes 120, 122 are printed on the upper surface 112 using conductive ink. The conductive ink can be printed on the top surface 112 by gravure printing, flexographic printing, screen printing, rotary screen printing, ink jet printing, ^or any other suitable printing process. The third electrode 124 is not shown in this illustration, but it should be understood that similar printing processes can be used to print the third electrode 124 on the upper surface 112 of the paper substrate.

In the illustrated example of FIG. 2C, the first and second tracks 126, 130 and the first and second contacts 128, 132 are disposed on the upper surface 112 of the paper substrate 102. In some examples, the first and second tracks 124, 128 and the first and second contacts 126, 130 are deposited on the top surface 112 using conductive inks through a printing process such as, for example, intaglio printing, flexographic printing, screen printing, rotary screen printing ^or ink jet printing. In some examples, the electrodes 120, 122 (and / or 124), I ^will traces 126, 130 (and / or 134) and contacts 128, 132 (and / or 136) are printed on the paper substrate 102 for a only printing process. In other examples, each of ^the electrodes, traces, and / or contacts can be printed through a separate printing process (for example, the first electrode 120 is printed through a first rotary screen printer and the second electrode 122 is printed through a second rotary screen printer).

As illustrated in FIG. Illustrative 2D, one or more reagents 200 are disposed on top of the first electrode 120. As the fluid sample comes into contact with the first electrode 120, the fluid sample mixes with the reagent 200 and produces biomolecular interaction. The reagent 200 may be deposited onto the surface (eg, the top surface and / or the side surface (s) of the first electrode 120 through a printing process such as, for example, ink jet printing, flexographic printing, gravure printing, screen printing, ^or slot die deposition. In other examples, reagent 200 is embedded in a hydrogel ^or other polymer and printed as a suspension, which is then cured (for example, by ultraviolet light ( UV) or electropolymerization) In other examples, the reagent 200 is mixed (eg, functionalized) directly into the material of the first electrode 120. In that case, as the fluid sample comes into contact with the first electrode 120 , a reaction occurs on the surface of the first electrode 120 that can be measured (eg, correlated) as the presence, absence, and / or concentration of the target analyte.

In the illustrated example of FIG. 2E, the hydrophilic ink layer 138 is deposited on the upper surface 112 of the paper substrate 102 on top of the first and second electrodes 120, 118, and the hydrophilic ink channel 140 is deposited on the upper surface 112 of the paper substrate 102. In some examples, the hydrophilic ink in layer 138 and the hydrophilic ink in channel 140 are the same ink. In other examples, different ink can be used to affect different flow rates through channel 140 and layer 138, as previously disclosed. In operation, a fluid sample may be deposited at the first end 142 of channel 140, and the hydrophilic ink in channel 140 picks up (e.g., by capillary action) the fluid sample toward layer 138, which covers sensor 118 ( FIG. 1). As a result, the sample contacts and interacts with reagent 200, which produces an electrical signal that is detectable by first electrode 120. Hydrophilic ink from channel 140 and / or hydrophilic ink layer 138 are deposited on substrate 102 through one of the processes disclosed above.

In the illustrated example of FIG. 2F, seal layer 148 is deposited over layer 138 and channel 140. Seal layer 146 is a fluid impermeable layer, which reduces the amount of evaporation from the sample and the risk of contamination such as, for example , produced by accidentally touching the hydrophilic ink by an operator. In the illustrated example, the first end 142 of channel 140 is not covered by seal layer 148, so that the fluid sample can be deposited on the hydrophilic ink of channel 140.

FIG. 3A is a cross-sectional view of illustrative device 100 taken along line BB of FIG. 1. In the illustrated example, the sealing layer 148 covers the top and I side ^I of the channel 140 and therefore, there is a free leak path formed or defined by the seal layer 148 (for example, the upper limit ) and the upper surface 112 of the paper substrate 102 (eg, the lower limit). Detection zone 116 is shown remotely behind channel 140.

FIG. 3B is a cross-sectional view of illustrative device 100 taken along line CC of FIG. 1. In the illustrated example, the first electrode 120 is disposed on the upper surface 112 of the paper substrate 102. In some examples, the reagent 200 is disposed on the upper surface of the first electrode 120. The hydrophilic ink layer 138 of the detection zone 116 covers the electrode 120 and the surrounding area so that a fluid sample can be mixed with the reagent and contact the first electrode 120. the sealing layer 148 covers the top and sides of the I ^os layer 138 of hydrophilic ink and thus creates a leak-free area that is defined by the sealing layer 148 (eg, the upper limit) and the upper surface 112 of the paper substrate 102 (eg, the lower limit). Also illustrated are the second and third traces 130, 134, which they are connected to the second and third contacts 132, 136, respectively (FIG 1).

An illustrative 400 for use with the illustrative substrate diagnostic device 100 is illustrated in FIG. 4. After a sample is deposited at the first end 142 of channel 140, absorbed through channel 140 and layer 138 into detection zone 116, and mixed with reagent 200, the electrical signals produced by the reaction can be read. To read the signals, the second end 106 of the sheet of paper 102 can be inserted into a slot 402 of the reader 400. The slot includes a plurality (eg, three) of pins (or contact points) that are connected to the first, second, and third contacts 128, 132, 136. In the illustrated example, reader 400 includes a screen 404 (eg, a user interface, a display screen, etc.) and a plurality of buttons 406 for interacting with screen 404. In the illustrated example, reader 400 detects the analyte of interest using, for example, amperometric, voltammetric, and / or potentiometric techniques. For example, reader 400 can measure (eg, through a processor) the voltage difference between the first electrode pair 120 and the second electrode pair 122 (eg, the working electrode / counter electrode pair) and the third electrode pair 124 and the second electrode pair 1222 (eg, the reference electrode / counter electrode pair). The difference in voltage between the two pairs can be correlated with the presence, absence and / or concentration of the target analyte. In some examples, device 100 includes a plurality of electrodes that form different sensors to detect analytes in the sample. In such an example, the reader may include additional pins to connect the additional contacts, and the reader 400 may perform different measurements sequentially and / or simultaneously.

FIG. 5 is a diagram of an illustrative system ^or assembly 500 for creating (eg, manufacturing, fabricating, constructing) a substrate diagnostic device such as, for example, the illustrative device 100 of FIG.

1. Set 500 includes a series or plurality of rollers, including a first roller 502 and a second roller 504, that operate in synchronized rotation to drive a substrate 506 through set 500. In some examples, set 500 includes rollers to move base substrate 506 through the array using continuous roll-fed ^or roll-to-roll processing techniques. Other examples can use conveyors, pulleys and / or any other suitable transport mechanism.

In illustrative assembly 500, first roller 502 rotates to unwind substrate 506, which, in some examples, is a single sheet in a rolled configuration. In the illustrated example, substrate 506 is a hydrophobic (eg, fluid impervious) sheet of paper such as, for example, paper substrate 102 (FIG. 1 and 2A). In some examples, substrate 506 is made from materials (eg, tree pulp) and / or chemicals that result in a hydrophobic paper substrate. In other examples, substrate 506 starts out as traditional paper and is then treated with wax ^or additional coatings / layers to create a hydrophobic surface. In some examples, substrate 506 is a continuous roll of hydrophobic paper. However, in other examples, substrate 506 may be punched, punched, ^or sawed into defined ^or distinct sheets ^or strips (eg, which are then converted into individual diagnostic devices).

In the illustrated example of FIG. 5, the assembly 500 includes a first printing station 508 and a second printing station 510 for printing electrodes (eg, electrodes 120, 122, 124) onto substrate 506 (eg, onto the top surface of substrate 506) . In the illustrated example, the first print station 508 prints a first type of electrode (eg, having Ag / AgCl) on the substrate 506, and the second print station 510 prints a second type of electrode (eg, having has Au) on substrate 506. In some examples, two or three electrodes are printed on substrate 506. For example, a first electrode can be a working electrode made of A ^u and a second electrode can be a counter electrode made of A ^u . Also, a third electrode can be a reference electrode made of Ag / AgCl. In such a case, the first print station 508 may include A ^u ink to print the first and second electrodes and the second print station 510 may include Ag / AgCl ink to print the third electrode. In other examples, the first and second print stations 508, 510 are interchanged.

In the illustrated example, the first and second printing stations 508, 510 are rotary screen printers. In general, a rotary screen printer uses a screen that has an image (eg, a negative of the shape) curved around a cylinder. As the image on the screen rotates to contact a substrate, an ink passes through the holes in the screen and is printed on the substrate. The screens of the rotary screen printers 508, 510 can include the shapes of the electrodes to be deposited on the base substrate 506. As the cylinders rotate, the ink fills the negative image on the screen and the image (for For example, the electrode (s) are printed on the base substrate 506. For example, the device 100 of FIG. 1 includes the first, second, and third electrodes 120, 122, 124, which are deposited on the top surface 112 of the paper substrate 102. An illustrative cross-sectional view of the paper substrate 102 having the electrodes is illustrated in FIG 2B.

In some examples, the first and second print stations 508, 510 also print traces (eg, wires, cables) and contacts on the substrate 506. The contacts can be used to communicatively couple the electrodes to a reader, as previously disclosed. . For example, device 100 of FIG. 1 includes the first, second, and third contacts 128, 132, 136, and the first, second, and third traces 126, 130, 134, and an illustrative cross-sectional view of device 100 at said printing stage is illustrated in FIG. 2 C.

In some examples, only one electrode is Printed on substrate 506. In other examples, more than one electrode is Printed on substrate 506. In some examples, all electrodes are Printed on substrate 506 at the same Print station. In other examples, multiple Print Stations are used to Print multiple electrodes and / or produce electrodes with multiple layers.

In some examples, a reagent is deposited on one ^or more of the electrodes through an optional third Print Station 512 (shown in dashed lines). The third Print Station 512 employs an Inkjet Printer to deposit one ^or more reagents. In other examples, the reagents are Printed directly onto the top surface of substrate 506 (eg, near the electrode (s)). For example, device 100 includes reagent layer 200 (FIG. 2D) that is disposed on top of first electrode 120 (eg, working / active electrode). Reagent 200 is illustrated in cross-sectional views of FIGS. 2D-2F.

In Illustrative assembly 500 of FIG. 5, a hydrophilic ink or slurry material layer (eg, layer 138) and a hydrophilic ink ^or slurry material channel (eg, channel 140) are deposited onto substrate 506 in a fourth Print station 514 The ink used in the layer and channel can include a suspension of hydrophilic particles (eg, microbeads or nanobeads) mixed with a binder. The layer is imprinted on the electrodes (and the reagent) and on the surface of the substrate 506 between the electrodes. In the Illustrated example, the fourth Printing station 514 is a Rotary Screen Printer. The screen of the Rotary Screen Printer can include a negative image of the channel and the layer to be printed on the substrate 506.

In some examples, the channel and layer of assembly 500 are printed on substrate 506 at the same time. In other examples, the assembly 500 includes multiple Print stations, and the channel and layer can be Printed separately and / or by multiple Print stages.

In the Illustrated example, a seal layer (eg, seal layer 148 of FIG. 1) is deposited over the layer and channel in a fifth Print station 516. The seal layer is a hydrophobic ink, which creates a Fluid Impervious layer over the layer and the hydrophilic ink channel. In some examples, the sealing layer is only deposited on the hydrophilic layer and the channel, and is not deposited on the remainder of the substrate 506. As a result, less hydrophobic ink is used and therefore reduces manufacturing cost.

In some examples, the base substrate 506 is diced, ^or cut ^or otherwise separated into individual devices at an optional cutting station 518 (shown in dashed lines). In other examples, the cutter station 518 only creates a groove between two devices. In this case, the devices can be separated later and / or in a different Installation.

The Illustrative set 500 of the Illustrated example uses a reel-to-reel (R2R) process (eg, continuous roll-fed, roll-to-roll processing) where substrate 506 is unwound from first roll 502 and wound onto second roll 504 at the opposite end of the process. However, in other examples, other types of Printing processes can be used.

FIG. 6 is a block diagram of an Illustrative processing system 600 for use with a substrate diagnostic device build assembly such as, for example, assembly 500 of FIG. 5 to create, for example, the paper device 100 of FIG. 1. Illustrative processing system 600 includes a controller 602, which controls the operation of assembly 500 through selected controller components.

For example, the processing system 600 includes a roller controller 604, which controls one or more rollers (eg, the first and second rollers 502, 504) of the set 500. In some examples, the processing system 600 includes one or more plus 604 roller drivers. In the example shown, the roller controller (s) 604 are communicatively coupled to one or more rollers 606a-n. Rollers 606a-n may correspond to, for example, the first and second rollers 502, 504 of the Illustrative set 500. In some examples, additional rollers (eg, the rollers from the Individual Print Stations) are used in the assembly, and the roller controller (s) 604 controls the respective rollers 606a-n. The roller controller (s) 604 control the rotation of the rollers 606a-n using, for example, a motor, to regulate one ^or more operating characteristics of the rollers. Such operating characteristics may include the speed of rotation, duration of rotation, direction of rotation, acceleration, etc. of rollers 606a-n. For example, the rotation speed can be used to determine how long a portion of the substrate is exposed to one ^or more Print stations (eg, one or more of Print stations 508-514). Thus, roller controllers 604 control the speed at which one or more substrates are processed. In the Illustrated example, a processor 608 operates the roll controller (s) 604 and thus rolls 606a-n in accordance with a roll protocol.

In the Illustrated example of FIG. 6, the processing system 600 Includes a Printer controller 610 that controls one or more of the Printers in the set 500. In some examples, the Illustrative processing system 600 Includes one or more Printer controllers 610. In the example shown, the Printer driver (s) 610 are communicatively coupled to one ^or more Printers 612a-n. The 612a-n Printers can correspond, for example, to Printers 508-514 of the Illustrative set 500. The printer controller / s 612a-n control, for example, the thickness, width and / or pattern of ^the electrode / s, ^the contacts), the trace / s, the hydrophilic channel , the hydrophilic layer and / or the sealing layer material applied to the substrate 506 by the respective printing stations 508-514. In the examples where the ink (for example, the conductive ink of ^the electrode (s), the hydrophilic ink of the layer and the channel, the hydrophobic ink of the sealing layer) is applied by rotary screen printing, ^the controllers Printer 610 can control the pressure of ^the rollers associated with the respective printers 508, 510, 514, 516 and thus affect the quality of the ink applied to the substrate. In some examples, the 612a-n printers operate in connection with the 606a-n rollers. In these examples, the printer drivers 610 work in association with the roller drivers 604 to define, for example, a rate at which the electrode (s), channel, layer, and sealing layer are deposited on the substratum. In the illustrated example, processor 608 operates printer controller (s) 610 and thus printers 612a-n according to a conductive ink, hydrophilic ink, and / or hydrophobic ink application protocol.

Illustrative processing system 600 also includes a cutting station controller 614 that controls a cutting station 616. In some examples, illustrative processing system 600 includes one ^or more cutting station controllers 614. In the example shown, the controller (s) 614 of the cutting station are communicatively coupled to one ^or more cutting stations 616. The cutting station (s) 616 may correspond to, for example, the cutting station 518 from the illustrative set 500. EI / I ^I drivers cutting station 614 controlling, for example, cutting ^or dividing I ^I substrates (for example, I ^will diagnostic devices on substrate individual paper), a size of the discrete units in that ^the substrates are cut, a gap between discrete units formed from continuous substrates, an operating speed of a cutting instrument, the retraction of the cutting instrument, etc. In the illustrated example, the processor 608 operates the cutting station controller (s) 614 and thus the cutting station (s) 616 in accordance with a substrate cutting protocol.

In the illustrated example of FIG. 6, the processing system 600 includes a database 618 that can store information relating to, for example, the operation of the illustrative system 500. The information may include, for example, information on the length and dimensions of the substrates to be fed through the array 500; The materials and patterns of the electrodes, the contacts, the traces, the hydrophilic layers, the hydrophilic channels, the sealing layers and / or ^the reagents; the rotational characteristics of ^the rollers, such as a speed and / or a diameter; the properties of the material / s conductive, hydrophobic, hydrophilic / s, adhesive / s and / or other material / s to be applied to the substrates, etc.

The example of the processing system 600 of FIG. 6 includes a user interface such as, for example, a Graphical User Interface (GUI) 620. An operator or technician interacts with the processing system 600 and thus the illustrative assembly 500 via the interface 620 to provide, for example, commands related to the operation of ^the rollers 606a-n, such as speed, duration of rotation, etc. of the rollers; the pattern / s to be deposited through printers 612a-n; materials and patterns of the electrodes, the contacts, the traces, the hydrophilic layers, the hydrophilic channels, the sealing layers and / or ^the reagents; the size of the discrete units into which ^the substrates are cut through the cutting station 616; etc. Interface 620 can also be used by the operator to obtain information regarding the status of any substrate processing completed and / or in progress, to verify parameters such as speed and alignment, and / or to perform calibrations.

In the illustrated example, ^the components 604, 608, 610, 614, 618 of the processing system are communicatively coupled to other components of the illustrative processing system 600 via communication links 622. The communication links 622 can be any type. wired connection (for example, a data bus, a USB connection, etc.) and / or any type of wireless communication (for example, radio frequency, infrared, etc.) using any past, present or future communication protocol (for example, Bluetooth, USB 2.0, USB 3.0, etc.). Also, ^the components of the illustrative system 600 can be integrated into one device or distributed across two or more devices.

FIG. 7 illustrates another example of a substrate diagnostic device 700, which is not in accordance with the present invention (eg, a paper-based detection device) that can be used for AI ^or CC testing. Similar to previously disclosed device 100, device 700 includes a detection zone, a channel, and a sealing layer. However, in device 700, the detection zone and channel are defined within the substrate material, which is hydrophilic, by hydrophobic ink barriers printed on the substrate.

For example, in the illustrated example of FIG. 7, device 700 includes a substrate 702, which is implemented as a hydrophilic sheet of paper. In other words, the substrate 702 is porous and is capable of absorbing and / or retaining liquids. In the illustrated example, the paper substrate 702 has a first end 704, a second end 706 opposite the first end 704, a first side 708, a second side 710 opposite the first side 708, a top surface 712, and a bottom surface 714 opposite top surface 712. In the illustrated example, the paper substrate 702 is substantially rectangular. However, in other examples, the paper substrate 702 may have another shape such as, for example, a square, a triangle, a circle, an ellipse, an irregular shape, and so on.

In the Illustrated example, device 700 includes a detection zone 716 (eg, a test area, a reaction zone, etc.) that defines a location where the test will be performed. Detection zone 7-16 has a sensor 718 to detect a reaction between the sample and a reagent, which is then used to measure the presence, absence, and / or concentration of the target analyte in the sample. In the illustrated example, sensor 718 includes a first electrode 720 (for example, the working electrode), a second electrode 722 (for example, a counter electrode, a ground electrode), and a third electrode 724 (for example, a ground electrode). reference). The first, second, and third electrodes 720, 722, 724 may function in a similar manner to the first, second, and third electrodes 120, 122, 124 of illustrative device 100. In the illustrated example, however, the first, second and third electrodes 720, 722, 724 are printed on the material of the substrate 702. Because the substrate 702 is hydrophilic, the conductive ink of the first, second and third electrodes 720, 722, 724 is absorbed into the substrate material 702. In some examples, the one ^or more of the electrodes 720, 722, 724 are made of gold (Au), gold on silver chloride (AgCl), carbon (C), and / or silver / silver chloride (Ag / AgCl). The one or more of the electrodes 720, 722, 724 can be printed on the substrate 702 using, for example, gravure printing, flexographic printing, screen printing, rotary screen printing and / or ink jet printing. In other examples, other suitable types of printing processes may be employed to print the conductive ink onto the 702 paper substrate.

In the illustrated example of FIG. 7, device 700 includes a first trace 726 that electrically connects the first electrode 720 to a first contact 728, a second trace 730 that electrically connects the second electrode 722 to a second contact 732, and a third trace 734 that electrically connects the third electrode 724 to a third contact 736. Device 700 can be used by an electrical reader (eg, reader 400), and contacts 728, 732, 736 allow the reader to electrically communicate with electrodes 72nd, 722, 724 and detect electrical signals including, for example, changes in, for example, amps, volts, and / or resistance.

To create a boundary for the detection zone 716 and / or a channel to capture the sample in the detection zone 716, the device 700 includes a hydrophobic barrier 738. The hydrophobic barrier 738 is a hydrophobic ink line that is printed on the substrate 702 and forms ^or defines a pathway in between. To allow the sample to contact electrodes 720, 722, 724 on substrate 702, a layer 740 of hydrophilic mesh (eg, hydrophilic material suspension ink) is deposited on substrate 702 in detection zone 716 The hydrophilic mesh is absorbed into the paper substrate material 702. The hydrophilic mesh may include hydrophilic ink similar to that of layer 138 disclosed above. The hydrophilic mesh has increased the uptake ability to capture a sample through the detection zone 716. Furthermore, in the illustrated example, a channel 742 of the hydrophilic mesh is deposited in the paper substrate material 702 to form a channel from the first end 704 of the paper substrate 702 to the layer 740. The hydrophilic mesh is absorbed into the material of the paper substrate but is limited by the hydrophobic barrier 738. In other words, the hydrophilic mesh can be printed on the paper substrate 702, and hydrophobic barrier 738 prevents hydrophilic ink from spreading beyond hydrophobic barrier 738. The hydrophilic mesh of layer 740 and channel 742 may include hydrophilic ink similar to that of layer 138 and channel 140 disclosed above. relative to device 100. The hydrophobic mesh or layer 740 and / or channel 742 can be printed onto substrate 702 using a printing process such as, for example, intaglio printing, i flexographic printing, screen printing, rotary screen printing and / or ink jet printing.

FIG. 8 is a cross-sectional view of illustrative device 700 taken on line DD of FIG. 7. In the illustrated example, the hydrophobic barrier 738 is printed on the paper substrate 702 and absorbed into the material of the paper substrate 702. The hydrophobic barrier 738 repels ^or resists liquid and thus creates a pathway between the lines of the hydrophobic barrier 738. In particular, when the hydrophilic mesh is printed on the paper substrate, the hydrophilic ink is absorbed into the material of the paper substrate but is limited between the hydrophobic barrier 738. When a fluid sample is deposited (eg, pipetting) into an inlet 744 (FIG. 7) of channel 742, the sample is captured (eg, by capillary action) through the hydrophilic mesh (eg, the porous cellulose matrix) of the channel 142, between the lines of the hydrophobic barrier 738, and within the hydrophilic mesh of the layer 740. In this way, the sample is transferred through the channel 742 and to the detection zone 716 where the sample comes into contact with or wet the electrodes 720, 722, 724.

In some examples, the hydrophobic ink in the hydrophobic barrier 738 is printed using a printing process such as, for example, gravure printing, flexographic printing, screen printing, rotary screen printing and / or ink jet printing. In other examples, other suitable types of printing processes may be employed to print the hydrophobic ink on the substrate 702. In some examples, the hydrophobic ink may be printed on the substrate prior to printing the sensor conductive inks. For example, ink ^or hydrophobic barrier can be used to create a pattern to print electrodes 720, 722, 724, traces 726, 730, 734 and / or contacts 728, 732, 736. In some of these examples, this produces increased adhesion of conductive ink to substrate 702.

In the illustrated example of FIG. 8, the hydrophobic barrier 738 is only partially disposed in the thickness of the substrate 702. However, in other examples, the hydrophobic barrier 738 is completely absorbed into the substrate 702. In both examples, the hydrophobic barrier 738 creates a barrier between two sections substrate 702.

In some examples, one ^or more reagents are printed on the paper substrate 702. For example, a reagent may be printed on the substrate 702 in the detection zone 716. As the sample is captured in the detection zone 716, the sample interacts with the reagent, and the sensor 718 can detect biomolecular interaction.

Additionally or alternatively, one or more reagents may be printed on the first electrode 720. The reagent / s may be printed using, for example, in-line inkjet printing ^or high performance printing (e.g., printing up to 120 meters / minute) such as, for example, flexographic printing, screen printing or deposition on slotted dies. In some examples, the reagent (s) can / are embedded in a hydrogel or other polymer / s printed as a suspension which is then cured (eg, via ultraviolet light ^or electropolymerization). In some examples, a plurality of electrodes are used to perform different types of assays and the electrodes can be functionalized with different binding reagents to detect a plurality of analytes.

In some examples, a sealing layer 746 is applied to the top surface 712 of the paper substrate 702 to prevent evaporation of the sample and / or reduce the risk of contamination. Seal layer 746 creates a fluid impervious (or hydrophobic) surface on top of the hydrophilic mesh of channel 742 and detection zone layer 740 716. In some examples, seal layer 746 is a substrate hydrophobic (eg, a hydrophobic paper sheet) that is coupled (eg, laminated, bonded) to the top surface 712 of the paper substrate 702. In some examples, the sealing layer 746 is not disposed above the inlet 744 of the channel 742, so that the fluid sample can be deposited at the inlet 744 of the channel 742. In other examples, the entire channel 742 is covered with the layer. seal, and an opening ^or hole may be defined in the seal layer to allow the fluid sample to contact the substrate 702 at the inlet 744 of the channel 742.

In the illustrated example, hydrophilic mesh ^or ink is printed on substrate 702 to form channel 142 and layer 740. However, in other examples, a hydrophilic mesh is not used. Instead, the porous structure of substrate 702 itself forms channel 742 and layer 740. For example, a fluid can be drawn through the porous / hydrophilic structure of substrate 702 between barrier 738.

In some examples, the composition of the conductive ink / s, hydrophobic ink / s, hydrophilic ink / s and / or dielectric ink / s) may depend on the properties of the substrate 702 and / I or ^I used print processes. For example, the conductive ink may need to be cured or dried after deposition (eg, via ultraviolet light, heat, infrared (IR), or air drying) depending on the ink composition and / or substrate tolerances.

FIG. 9 is a diagram of an illustrative system or assembly 900 for creating (eg, manufacturing, producing, creating, constructing, fabricating) a paper substrate diagnostic device such as, for example, device 700 of FIG. 7. Assembly 900 includes a series ^or plurality of rollers, including a first roller 902 and a second roller 904, that operate in synchronized rotation to drive a substrate 906 through assembly 900. In some examples, assembly 900 includes rollers. to move substrate 906 through the array using continuous roll-fed or roll-to-roll processing techniques. Other examples can use conveyors, pulleys and / or any other suitable transport mechanism. In the illustrated example, a plurality of printing processes are provided to create one or more devices on substrate 906 so that ^the devices can be mass produced.

In illustrative assembly 900, first roller 902 rotates to unwind substrate 906, which, in some examples, is a single sheet in a rolled configuration. In the illustrated example, substrate 906 is paper and is hydrophilic (eg, it has a strong affinity for liquid). In some examples, substrate 906 is a continuous roll of hydrophilic paper. However, in other examples, the substrate 906 may be cut, sawn, or otherwise separated into defined ^or distinct sheets ^or strips (eg, which are then converted into individual diagnostic devices). Substrate 906 may correspond, for example, to paper substrate 702 of device 700. An illustrative cross-sectional view of paper substrate 702 is illustrated in FIG. 8.

In the illustrated example, which is not in accordance with the present invention, assembly 900 includes a first printing station 908 for printing a hydrophobic ink barrier on substrate 906. Hydrophobic ink is printed on substrate 906 to define a pathway and a detection zone for testing. Once dry, the hydrophobic ink creates a barrier (for example, the 738 barrier) that prevents a sample from being captured ^or absorbed onto the substrate in areas outside the defined detection pathway and area. In the illustrated example, print station 908 is a flexographic printer. In general, flexographic printing involves the use of a relief plate. In the illustrated example, the plate is wrapped around a roller, so that as the roller rotates, it deposits an image (eg, the lines that define the channel and the detection zone) on the substrate 906. In addition or alternatively, other types of printing processes can be implemented to print the hydrophobic ink such as, for example, intaglio printing, screen printing, rotary screen printing or ink jet printing.

In the illustrated example, the assembly 900 includes a second printing station 910 and a third printing station 912 for printing electrodes on the substrate 906. In the illustrated example, the second printing station 910 prints a first type of electrode (for example, having Ag / AgCl) on substrate 906, and the third print station 912 prints a second type of electrode (for example, having AuNP) on base substrate 906. In some examples, two ^or three electrodes are printed on substrate 906. For example, a first electrode can be a working electrode made of A ^or NP and a second electrode can be a counter electrode made of AuNP. Furthermore, a third electrode can be a reference electrode made of Ag / AgCI. In such a case, the third print station 912 may include AuNP ink to print the first and second electrodes and the second print station 910 may include Ag / AgCl ink to print the third electrode. In other examples, the second and third print stations 910, 912 are interchanged. In the illustrated example, the second print station 910 is a rotary screen printer and the third print station 912 is an inkjet print. However, in other examples, may implement other types of printing processes to print I ^will electrodes on the substrate 906. T ^he electrodes may correspond, for example, the first, second and third electrode 720, 722, 724 of the device 700 .

The conductive ink for ^the electrodes is deposited on the substrate 906 and absorbed into the material of the substrate 906. Thus, in the illustrated example, the electrodes are disposed at least partially within the material of the substrate 906. In some examples, the Second and third print stations 910, 912 also print traces (eg, wires, cables) and contacts on substrate 906. The contacts can be used to connect the diagnostic device with a reader, so that the reader includes pins or contacts that match the device's contacts. For example, device 700 of FIG. 7 includes the first, second, and third contacts 728, 732, 736, and the first, second, and third traces 726, 730, 734.

In some examples, only one electrode is printed on the substrate 906. In other examples, more than one electrode is printed on the base substrate 906. In some examples, all the electrodes are printed on the substrate 906 at the same printing station. In other examples, multiple print stations are used to print multiple electrodes and / or produce multiple layer electrodes.

In the illustrated example, assembly 900 includes a sintering station 914 for sintering one ^or more of ^the electrodes. In general, sintering is a process to form a solid mass of material by heat and / or pressure. The sintering process causes ^the atoms of the material to diffuse through ^the limits of the particles, fusing the particles and creating a solid piece. Sintering station 914 may include a source of heat and / or pressure to sinter (eg, fuse) the material from each of the electrodes. Sintering station 914 can operate by applying alternating current or direct current to the electrode (s) to induce coalescence of the conductive material.

In the illustrated example, the assembly includes a fourth print station 916 for depositing one or more reagents on the electrodes or around the electrodes on the substrate 906. In the illustrated example, the fourth print station 916 is a inkjet printer. ink. In addition ^or alternatively, other types of printing processes may be implemented to deposit one or more reagents on top of substrate 906, such as, for example, flexographic printing, screen printing, or slot die deposition.

In some examples, a hydrophilic mesh layer is deposited (eg, printed) on substrate 906 in the channel (eg, channel 742) and detection zone (eg, detection zone 740) areas ) to increase the ability of the sample to be captured through substrate 906. In the illustrated example, assembly 900 includes a fifth mesh printing ^or laminating station 918 that deposits (eg, prints and fixes) a hydrophilic ink on the substrate 906. The hydrophilic ink passes through or is absorbed by the substrate material 906 between the hydrophobic barrier to create the channel and the detection zone. In some examples, the mesh laminating station 918 includes a printer such as, for example, a gravure printer, a flexographic printer, a screen printer, a rotary screen printer, and / or an ink jet printer.

In the illustrated example, a sealing layer (eg, sealing layer 746 of FIG. 7) is disposed over substrate 906 to seal the channel and detection zone to reduce the probability of sample evaporation. and / or contamination. In the illustrated example, assembly 900 includes a third roll 920 that unwinds a hydrophobic (eg, fluid impervious) substrate 922. Hydrophobic substrate 922 may be, for example, a hydrophobic sheet ^or piece of paper. Hydrophobic substrate 922 is laminated on top of substrate 906 to seal the channel and detection zone of the device. As a result, a substantially fluid-sealed pathway is created in substrate 906 between the hydrophobic barrier and the seal layer.

In the illustrated example, the assembly 900 includes a laser 924 (e.g., a CO2 laser, an excimer laser) to cut the hydrophobic substrate 922 into a specific pattern and / or create sample and / or reagent entry ports in the hydrophobic substrate 922. For example, laser 924 can change the hydrophobicity of hydrophobic substrate 924 in certain areas to create sample and / or reagent entry ports. The laser modifies the hydrophobicity of the hydrophobic substrate 922 and increases the ability of the hydrophobic substrate 922 to absorb and / or capture fluids (eg, samples and / or reagents). When the hydrophobic substrate 922 is coupled to the top of the substrate 906, the hydrophobic substrate 922 forms a waterproof plug to I ^os fluids on the device, and I ^os input ports sample and / or reagent allow the samples and / or I reagents ^will be deposited on the device (for example, through the hydrophobic substrate 922). In some examples, the degree of hydrophilicity can be modified by changing the extent of laser processing of the hydrophobic substrate. In this way, valve structures can be created to control the flow of samples and / or reagent within and / or through the device. In some examples, the hydrophobic substrate 906 is cut ^or shaped into a shape that matches that of the channel and the detection zone, so that only the channel and the detection zone are sealed. As a result, less material can be used when forming the seal layer.

In some examples, instead of using the hydrophobic substrate 922, a hydrophobic ink layer is printed on the substrate 906 to form the sealing layer. In some examples, the hydrophobic ink is only printed on top of the channel and detection zone.

In the illustrated example, the assembly 900 includes a lamination unit 926 for coupling the hydrophobic substrate 922 to the top of the substrate 906. The lamination unit 926 may include a source of heat and / or pressure to adhere the two substrates 906, 922 with each ^other . In some examples, adhesive (for example, a glue) can be used. In some examples, several additional sheets of waterproof paper can be used to create a multi-layer laminated device.

In some examples, substrate 906 is diced or otherwise separated into individual devices. The illustrative assembly 900 includes a cutting station 928 that cuts the substrate 906 at the individual devices. In other examples, cutter station 926 creates a groove between two devices. In this case, the devices can be separated later and / or in a different installation.

The set 900 in the illustrated example uses reel-to-reel (R2R) processing (e.g., continuous roll-fed processing, roll-to-roll process) where substrate 906 is unwound from first roll 902 and wound onto second roll 904 at the opposite end of the process. The substrate 906 of the second roll 904 has a plurality of microfluidic diagnostic devices printed on the substrate 906. The device may correspond, for example, to the device 700 of FIG. 7.

FIG. 10 is a block diagram of an illustrative processing system 1000 for use with a substrate diagnostic device build assembly such as, for example, assembly 900 of FIG. 9. Illustrative processing system 1000 includes a controller 1002, which controls the operation of assembly 900 through selected controller components.

For example, illustrative processing system 1000 includes a roller controller 1004, which controls one or more of the rollers (eg, the first, second, and / or third rollers 902, 904, 920) of set 900. In some examples , the processing system 1000 includes one ^or more roller controllers 1004. In the example shown, the roller controller (s) 1004 are communicatively coupled to rollers 1006a-n. Rollers 1006a-n may correspond to, for example, the first, second, and / or third rollers 902, 904, 920 of illustrative assembly 900. In some examples, additional rollers (eg, the rollers from individual print stations) are used in the assembly, and the roller controller (s) 1004 controls the respective rollers 1006an. EI / I ^I / controller 1004 is roll control rotation of the rollers 1006a-n using, for example, an engine, for regulating one ^or more operating characteristics I ^os rollers. Such operational characteristics may include the speed of rotation, duration of rotation, direction of rotation, acceleration, and so on. of the rollers 1006a-n. For example, the speed of rotation can be used to determine how long a portion of the substrate is exposed to one or more print stations (for example, one or more of print stations 908, 910, 912, 916). . Thus, roller controllers 1004 control the speed at which one or more substrates are processed. Also, an illustrative processor 1008 operates the roller controller (s) 1004.

Illustrative processing system 1000 includes a printer controller 1010 that controls one ^or more of the printers in array 900. In some examples, illustrative processing system 10 includes one or more printer ^or 101 controllers. In the example shown, the printer driver (s) 1010 are communicatively coupled to printers 1012a-n. Printers 1012a-n may correspond, for example, to printers 908, 91 ^or , 912, 916 of illustrative set 900. EI / I ^I / controller is 1012a-n printer control, for example, the thickness, width and / or pattern of the / R ^I electrode / s, contact / s, trace / s, hydrophilic barrier and / or reactive / s applied to the substrate 906 by the printing stations 908, 91o, 912, 916. In the examples where the ink (for example, the conductive ink of ^the electrode / s, the hydrophobic ink of the hydrophobic barrier) is applied by rotary screen printer ^or flexographic printer, I ^os printer driver 1010 can control the pressure of Ios rollers associated with the respective printers 908, 910 and thus affect the quality of ink applied to the substrate. In some examples, printers 908, 91, ^or 912, 916 operate in connection with rollers 902, 904, 92. In these examples, the printer controller (s) 1010 work in association with the roller controller (s) 1004 to define, for example, a rate at which the electrode (s), reagent (s) , the hydrophobic barrier, etc. settle on the substrate. In the illustrated example, processor 1008 operates printer controller (s) 1010, and thus printers 1012a-n, according to a conductive ink, hydrophobic ink, and / or reactive ink application protocol.

Illustrative processing system 1000 includes a sintering station controller 1014 that controls the sintering station of illustrative assembly 900. In some examples, the processing system 1000 includes one ^or more sintering station controllers 1014. In the illustrated example, the one ^or more sintering station controllers 1014 are communicatively coupled to one or more sintering stations 1016. The sintering station (s) 1016 may correspond, for example, to the sintering station 914 of the assembly. 900 illustrative. EI / Ios 1014 controller of the sintering station can control, for example, the intensity of the pressure and / or heat applied to the substrate, the size of an area of the substrate exposed to pressure and / or heat, duration of exposure to pressure and / or heat, etc. In the illustrated example, the processor 1008 operates the controller (s) 1014 of the sintering station and, therefore, the sintering station (s) 1016 according to the application protocol of conductive ink and / or dielectric ink. .

Illustrative processing system 1000 includes a mesh lamination controller 1018 that controls the mesh laminating unit of illustrative assembly 900. In some examples, the processing system 1000 includes one or more mesh lamination controllers 1018. In the illustrated example, the one or more mesh lamination controllers 1018 are communicatively coupled to one or more mesh lamination units 1020. The mesh rolling units 1020 may correspond, for example, to the mesh rolling unit 918 of the illustrative assembly 900. EI / I ^I / controller is 1,018 laminating mesh can control, for example, the thickness, width and / or pattern of the ink layer or mesh hydrophilic applied to the substrate 906 by the lamination unit 918 mesh. In some examples, the mesh laminating unit 1020 is implemented as a printer and can be controlled, for example, by means of printer controller / s 1010. In the illustrated example, the processor 1008 operates the mesh laminating controller (s) 1018 and, therefore, the mesh laminating unit (s) 1020 according to the application protocol of a mesh layer and / or hydrophilic ink.

Illustrative processing system 1000 includes a laser controller 1022. In some examples, the illustrative processing system 1000 includes one ^or more laser controllers 1022. In the example shown, the one ^or more laser controllers 1022 are communicatively coupled to one ^or more lasers 1024 to control the laser (s) 1024. The laser (s) 1024 may correspond to, for example, the laser 924 of the Illustrative 900 set. The 1022 laser controllers control, for example, the intensity of the laser (s) is 1024, a size of the irradiation surface area with respect to the substrate (s), the depth at which the laser (s) is 1024 / n penetrates a substrate, a duration during which the laser (s) 1024 penetrate or not the substrate, and / or the extent to which the laser (s) 1024 affects / does not change the hydrophobicity of the substrate (eg, hydrophobic substrate 922). In the illustrated example, processor 1008 operates the laser controller (s) 1022 and thus the laser (s) 1024 according to a laser protocol.

Illustrative processing system 1000 includes a lamination unit controller 1026. In some examples, illustrative processing system 1000 includes one ^or more lamination unit controllers 1026. In the illustrated example, the one or more lamination unit controllers 1026 are communicatively coupled to one ^or more lamination units 1028 to control the lamination unit (s) 1028. The lamination unit (s) 1028 can / are correspond to, for example, lamination unit 926 of illustrative assembly 900. The controller / s of the laminating unit 1026 control, for example, the intensity of the pressure and / or heat applied to ^the substrate / s (for example, the hydrophobic substrate 922 and the substrate 906), the size of an area of ^the substrate / s exposed to pressure and / or heat, the duration of an exposure to pressure and / or heat, an amount of adhesive applied between the substrate / s, etc. In the illustrated example, processor 1008 operates lamination controller (s) 1026 and thus lamination unit (s) 1028 in accordance with a lamination protocol.

Illustrative processing system 1000 includes a cutting station controller 1030 that controls a cutting station 1032. In some examples, illustrative processing system 1000 includes one ^or more cutting station controllers 1030. In the example shown, the one ^or more cutting station controllers 1030 are communicatively coupled to one or more cutting stations 1032. The cutting station (s) 1032 may correspond to, for example, the cutting station 928 of the array. 900 illustrative. EI / I ^I drivers cutting station 1030 control, for example, cutting or splitting the / Ios substrates (for example, in devices Ios Diagnostic substrate individual paper), a size of the discrete units in that ^the substrates are cut, a gap between discrete units formed from continuous substrates, an operating speed of a cutting instrument, the retraction of the cutting instrument, etc. In the illustrated example, processor 1008 operates the cutting station controller (s) 1030 and thus the cutting station (s) 1032 in accordance with a substrate cutting protocol.

Illustrative processing system 1000 includes a database 1034 that may store information regarding, for example, the operation of illustrative system 1000. The information may include, for example, information on the length and dimensions of substrates (eg, substrate 906 and / or hydrophobic substrate 922) to be fed through assembly 900; I ^I I materials and patterns ^will electrodes, contacts I ^I, I ^I traces, hydrophobic barriers, the mesh layers ^or inks hydrophilic, the sealing layers and / or I ^I reagents; the rotational characteristics of the rollers, such as a speed and / or a diameter; the properties of the material / s conductive, hydrophobic, hydrophilic / s, adhesive / s and / or other material / s to be applied to the substrates, etc.

Illustrative processing system 1000 includes a user interface such as, for example, a Graphical User Interface (GUI) 1036. An operator ^or technician interacts with processing system 1000 and thus assembly 900 illustrative via interface 1036 to provide, for example, commands related to the operation of rollers 1006a-n, such as speed, duration of rotation, etc. of ^the rollers; the pattern / s to be deposited through printers 1012a-n; materials and patterns of the electrodes, the contacts, the traces, the hydrophilic layers, the hydrophilic channels, the sealing layers and / or ^the reagents; the size of the discrete units into which ^the substrates are cut through the cutting station 1032; etc. Interface 1036 can also be used by the operator to obtain information regarding the status of any substrate processing completed and / or in progress, to verify parameters such as speed and alignment, and / or to perform calibrations.

In the example shown, the components 1004, 1008, 1010, 1014, 1018, 1022, 1026, 1030, 1034 of the processing system are communicatively coupled to other components of the illustrative processing system 1000 through communication links 1038 ^. 1038 communication links can be any type of wired connection (for example, a data bus, a USB connection, etc.) and / or any type of wireless communication (for example, radio frequency, infrared, etc.) using any protocol of past, present, or future communication (for example, Bluetooth, USB 2.0, USB 3.0, etc.). Also, the components of the illustrative system 1000 can be integrated into one device ^or distributed across two ^or more devices.

FIG. 11 illustrates another illustrative substrate diagnostic device 1100 (eg, a paper detection device) that can be used for AI or CC testing. Similar to previously disclosed devices 100 and 700, device 1100 includes a detection zone, channel, and / or sealing layer. However, in device 1100, the detection zone and channel are defined within an etched channel formed on an upper surface of a hydrophobic substrate.

In the illustrated example of FIG. 11, device 1100 includes substrate 1102. In the illustrated example, substrate 1102 is implemented as a piece (eg, a strip, a sheet, a piece, etc.) of hydrophobic paper. Paper substrate 1102 is fluid impervious and repels fluids or substantially reduces the amount of fluid that is permeable to paper substrate 1102. In some examples, paper substrate 1102 is constructed of hydrophobic material (eg, from trees and / or chemicals with hydrophobic properties). In other examples, the paper substrate 1102 is a conventional piece of paper (eg, having pores and fibers) and has a hydrophobic coating that prevents fluids from penetrating the porous structure of the paper substrate 1102.

In the illustrated example, the paper substrate 1102 has a first end 1104, a second end 1106 opposite the first end 1104, a first side 1108, a second side 1110 opposite the first side 1108, a top surface 1112, and a bottom surface 1114 opposite top surface 1112. In the illustrated example, the paper substrate 1102 has a substantially rectangular shape. However, in other examples, the paper substrate 1102 may have another shape such as, for example, a square, a triangle, a circle, an ellipse, an irregular shape, etc.

In the illustrated example, the paper substrate 1102 includes a recess ^or recess 1116 formed within the upper surface 1112 of the paper substrate 1102. The recess 1116 defines an area where the channel and the detection zone (which are discloses in detail below). In this way, the channel and detection zone are relatively flush with ^or even flush with the upper surface 1112 of the paper substrate 1102. In some examples, the gap 1116 is etched (for example, via a laser ^or other suitable medium) on the top surface 1112 of the paper substrate 1102.

In the illustrated example, device 1100 includes a detection zone 1118 (eg, a test area, a reaction zone, etc.) that defines a location where the test will be performed. The detection zone 1118 has a sensor 1120 to detect a reaction between the sample and one ^or more reagents, which is then used to measure the presence, absence, and / or concentration of the target analyte in the sample. In the illustrated example, sensor 1120 includes a first electrode 1122 (for example, the working electrode), a second electrode 1124 (for example, a counter electrode, a ground electrode), and a third electrode 1126 (for example, a ground electrode). reference). The first, second, and third electrodes 1122, 1124, 1126 may function similarly to the first, second, and third electrodes 120, 122, 124 of the illustrative device 100 disclosed above. In the illustrated example, however, the first, second, and third electrodes 1122, 1124, 1126 are imprinted in the gap 1116 formed in the upper surface 1112 of the paper substrate 1102. Because the paper substrate 1102 is hydrophobic, the conductive ink of the first, second and third electrodes 1122, 1124, 1126 does not penetrate the substrate material 1102. In some examples, the one or more of the electrodes 1122, 1124, 1126 are made of gold (Au), gold on silver chloride (AgCl), carbon (C) and / or silver / silver chloride (Ag / AgCl). The one or more of the electrodes 1122, 1124, 1126 can be printed on the substrate 1102 using, for example, gravure printing, flexographic printing, screen printing, rotary screen printing and / or ink jet printing. In other examples, other suitable types of printing processes may be employed to print the conductive ink on the 1102 paper substrate.

In the illustrated example, the device 1100 includes a first trace 1128 that electrically connects the first electrode 1222 to a first contact 1130, a second trace 1132 that electrically connects the second electrode 1124 to a second contact 1134, and a third trace 1136 that electrically connects the third electrode 1126 to a third contact 1138. The device 1100 can be used by an electrical reader (for example, the reader 400), and the contacts 1130, 1134, 1138 allow the reader to communicate electrically with ^the electrodes 1122, 1124, 1126 and detecting the electrical signal, including I ^will change, for example, amps, volts and / or resistance.

To allow a sample to contact sensor 1120 during the reaction, detection zone 1118 of the illustrated example includes a suspending layer of material 1140 (eg, hydrophilic ink) that is disposed within gap 1116 and on top of the sensor. 1120 and / or the area surrounding sensor 1120. Layer 1140 facilitates the transport of a fluid sample. The hydrophilic ink ^or material suspension may be similar to the disclosed hydrophilic ink for layer 138 of device 100 and may be deposited using the previously disclosed printing process.

To transfer or move a fluid sample to detection zone 1118, illustrative device 1100 includes a channel 1142 of the suspension of hydrophilic material ^or ink (eg, silica, porous material). In the illustrated example, the hydrophilic ink is disposed within the gap 1116. In this way, the hydrophilic ink in channel 1142 captures the fluid sample by capillary action from one end of channel 1142 to the other end of channel 1142. Specifically, in In the illustrated example, channel 1142 includes a sample deposit area ^or a first end 1l44 (eg, an inlet), an intermediate section 1146, and a second end 1148 (eg, an outlet). Second end 1148 is in contact with hydrophilic ink layer 140 of detection zone 1118. The hydrophilic ink ^or material suspension may be similar to the hydrophilic ink disclosed for channel 140 of device 100 and can be deposited using the process of print previously disclosed.

FIG. 12 is a cross-sectional view of device 1100 taken along line EE of FIG. 11. In the illustrated example, the gap 1116 is formed in the upper surface 1112 of the paper substrate 1102. The gap 1116 can have any shape ^or pattern that defines the geometry of the channel and the detection zone printed thereon. In the illustrated example, the hydrophilic ink from channel 1142 is printed in gap 1116 so that the top of channel 1142 is substantially flush with or even flush with the top surface 1112 of paper substrate 1102. In other examples, the Channel 1142 extends above or beyond the top surface 1112 of the paper substrate 1102.

In some examples, one or more reagents are printed in the gap 1116 and / or on one or more of the electrodes 1122, 1124, 1126 before the hydrophilic ink. As the sample is captured through channel 1142 and enters layer 1140 of detection zone 1118, the sample interacts with the reagent (s) and the biomolecular interaction can be detected by electrodes 1122, 1124, 1126. The reagent (s) can be printed using, for example, in-line inkjet printing ^or high performance printing such as, for example, flexographic printing, screen printing ^or slot die deposition. In some examples, the reagent (s) can be embedded in a hydrogel ^or other polymer (s) printed as a suspension which is then cured (eg, via ultraviolet light or electropolymerization). In some examples, a plurality of electrodes are used to perform different types of assays and ^the electrodes can be functionalized with different binding reagents to detect a plurality of analytes.

In some examples, a sealing layer 1150 is printed on top of channel 1142 and detection zone 1118 to prevent evaporation of the sample and / or reduce the risk of contamination. Seal layer 1150 creates a fluid impermeable (or hydrophobic) surface on top of the hydrophilic ink in channel 1142 and detection zone layer 1140 1118. In some examples, seal layer 1150 is not arranged above the inlet 1144 of the channel 1142, so that the fluid sample can be deposited at the inlet 1144 of the channel 1142. In other examples, the entire channel 1142 is covered with the sealing layer 1150, and an opening or a hole in seal layer to allow fluid sample to contact substrate 1102 at inlet 1144 of channel 1142.

The illustrative device 1100 illustrated in FIG. 11 can be created using an assembly similar to the illustrative assembly 500 disclosed above. It can further include a step of etching in the set to form the hole in the substrate before the / R process ^I / s printing.

Although in FIGS. 6 and 10, an illustrative way of implementing assembly 500 and / or assembly 900 of FIGS. 5 and 9, one or more of the elements, processes and / or devices illustrated in FIGS. 6 and 10 can be combined, divided, rearranged, omitted, deleted, and / or implemented in any other way. In addition, illustrative controller (s) 602, 1002, roller controller (s) 604, 1004, illustrative processor 608, 1008, illustrative printer controller (s) 610, 1010, and 1014 controller (s) illustrative sintering station controller (s), illustrative mesh rolling controller (s) 1018, illustrative laser controller (s) 1022, illustrative cutting station controller (s) 614, 1030 / s, the illustrative databases 618, 1034 and / or, more generally, the illustrative processing systems 600 and 1000 of FIGS. 6 and 10 can be implemented by hardware, software, firmware, and / or any combination of hardware, software, and / or firmware. Thus, for example, any of R ^os controllers 602, 1002 illustrative, the / the controller / is 604, 1004 roll, the processor 608, 1008 illustrate, the / the controller / is 610, 1010 illustratively / s printer, the 1014 illustrative sintering station controller (s), the 1018 illustrative mesh rolling controller (s), the 1022 illustrative laser controller (s), the 614 controller (s) Illustrative cutting station 1030 / s, illustrative databases 618, 1034 and / or, more generally, the illustrative processing systems 600 and 1000 of FIGS. 6 and 10 could be implemented by means of one or more analog or digital circuits, logic circuits, programmable processor (s), Application Specific Integrated Circuit (s) (ASIC), logic device (s) / Programmable s (PLD, Programmable Logic Device) and / or logic device / s programmable / s in the field (FPLD, Field Programmable Logic Device). When reading any of the apparatus ^or system claims of the present patent to cover a purely software and / or firmware implementation, at least one of the illustrative controllers 602, 1002, the roller controller (s) 604, 1004, the illustrative processor 608, 1008, illustrative printer controller (s) 610, 1010, illustrative sintering station controller (s) 10l4, illustrative mesh lamination controller (s) 1018 , the illustrative laser controller (s) 1022, the illustrative cutting station controller (s) 614, 1030, and / or the illustrative databases 618, 1034 are expressly defined to include a computer-readable tangible storage or storage disk such as a memory, a digital versatile disk (DVD, Digital Versatile Disk), a compact disk (CD, Compact Disk), a Blu-ray disc, etc. that store the software and / or firmware. In addition, the illustrative processing systems 600 and 1000 of FIGS. 6 and 10 may include one ^or more elements, processes and / or devices in addition to, or instead of, those illustrated in FIGS. 6 and 10 and / or may include more than any one or all of the illustrated process elements and devices.

In FIGS. 13 and 14, representative flowcharts of illustrative machine-readable instructions are shown for implementing the illustrative processing systems 600 and 1000 of FIGS. 6 and 10. In this example, the machine-readable instructions comprise a program for ITS execution by a processor such as processor 1512 shown in the illustrative processor platform 1500 described below in connection with FIG. 15. The program may be incorporated into software stored on a tangible, computer-readable storage medium such as a CD-ROM, floppy disk, hard disk, digital versatile disk (DVD), Bluray disk, or memory associated with processor 1512, but the entire program and / or parts of it could alternatively be executed by a device other than processor 1512 and / or embedded in dedicated firmware or hardware. Furthermore, although the illustrative program is described with reference to the flow diagram illustrated in FIGS. 13 and 14, alternatively, many other methods can be used to implement the illustrative processing systems 600 and 1000 of FIGS. 6 and 10. For example, the order of execution of the blocks can be modified and / or some of the described blocks can be modified, deleted ^or combined.

As mentioned above, the illustrative processes of FIGS. 13 and 14 can be implemented using encoded instructions (for example, computer-readable and / or machine-readable instructions) stored on a tangible computer-readable storage medium, such as a hard disk drive, a flash memory, a read-only memory ( ROM, Read-Only Memory), a compact disk (CD, Compact Disk), a versatile digital disk (DVD), a cache memory, a random access memory (RAM, Random-Access Memory) and / or any other storage device. storage or storage disk where information is stored for any duration (for example, for long periods of time, permanently, for short instances, for temporary storage and / or caching of information). As used herein, the term "computer-readable tangible storage medium" is expressly defined to include any type of computer-readable storage device and / or storage disk and to exclude propagation signals and to exclude transmission media. As used herein, "tangible computer-readable storage medium" and "tangible machine-readable storage medium" are used interchangeably. In addition or alternatively, the illustrative processes of FIGS. 13 and 14 can be implemented using encoded instructions (eg computer and / or machine readable instructions) stored on a non-transient computer and / or machine readable medium, such as a hard disk drive, flash memory, memory read-only, compact disc, digital versatile disc, cache memory, random access memory and / or any other storage device ^or storage disc where information is stored for any duration (for example, for long periods of time). time, permanently, for brief instances, for temporary storage and / or caching of information). As used herein, the term "non-transient computer-readable medium" is expressly defined to include any type of computer-readable storage device and / or storage disk and to exclude propagation signals and to exclude transmission media. As used herein, when the term "at least" is used as a transitional term in a preamble to a claim, it is open in the same way that the term "comprising" is open.

FIG. 13 depicts a representative illustrative flow diagram of an illustrative method 1300 for creating a substrate diagnostic device such as, for example, device 100 and / or device 1100, using, for example, assembly 500 illustrated in FIG. 5 and implemented / controlled by the processing system 600 illustrated in FIG. 6. Illustrative method 1300 includes unwinding a base substrate (block i 302). In the illustrated example, the base substrate is a hydrophobic (eg, fluid impervious, fluid resistant) sheet of paper (eg, a web of paper). In some examples, the base substrate is made from tree pulp and / or chemicals that produce hydrophobic paper. In other examples, the base substrate can be a traditional paper sheet that has been coated ^or treated with a hydrophobic layer ^or coating (eg, wax). The hydrophobic paper sheet may correspond, for example, to the paper substrate 102 of the 10o device, the base substrate 5O6 of the 5OO set and / or the paper substrate 1102 of the 11OO device. The 1O2 paper substrate, the 5O6 base substrate and / or the 1102 paper substrate provide a support surface for the sensor (s) and hydrophilic ink (s) to be printed on the substrate. Unwinding of the base substrate can be controlled, for example, by means of roller controller / s 6O4.

In some examples, the 13OO method includes determining if a hole ^or notch will be created in the surface of the base substrate (block 1304). A recess or recess provides an area where a channel and / or detection zone can be deposited (disclosed in detail below). The walls of the gap help shape the shape of the channel and / or the detection zone. If a gap is to be created in the base substrate, method 1300 includes etching a gap (block 1306). In some examples, the hole is etched using a laser. For example, device 1100 includes gap 1116 that forms ^or defines an area where channel 1142 and detection zone 1118 are disposed.

Illustrative method 1300 includes printing a working electrode and a counter electrode on the base substrate (block 1308). The working electrode and counter electrode form an electrical sensor that can be used to determine the presence, absence, and / or concentration of a target analyte in a sample. Different electrical techniques such as amperometric, voltammetric and / or potentiometric techniques can be implemented. The working electrode and the counter electrode can be printed on the base substrate (for example, on the upper surface of the base substrate or in a gap on the upper surface of the base substrate) through a rotary screen printer using, for example, inks conductive (for example, metallic). For example, in device 100 of FIG. 1, the first electrode 120 and the second electrode 122 are disposed on the upper surface 112 of the paper substrate 102. Similarly, in the device 1100 of ^f I ^g . 11, the first electrode 1122 and the second electrode 1224 are disposed within the gap 1116 in the paper substrate 1102. In the illustrative processing assembly of FIG. 5, the working electrodes and / or counter electrodes are printed on the base substrate 506 in the first and / or second printing stations 508, 510, which are implemented as rotary screen printers. The electrode printing stations may be controlled by, for example, the printer controller (s) 610 of the processing system 600 illustrated in FIG. 6. In other examples, other types of sensors can be printed on the base substrate. For example, some devices may include an optical or magnetic sensor, and the components thereof may be deposited on the base substrate (or within a gap in the base substrate).

Illustrative method 1300 includes determining if a reference electrode is to be used (block 1310). In some examples, a reference electrode is used, and the difference between the working electrode / counter electrode pair and the reference electrode / counter electrode pair is used to determine the change across the electrodes. The difference can be correlated with the change produced by the sample and the reagent and, therefore, is indicative of the presence, absence and / or concentration of the target analyte. If a reference electrode is to be added, illustrative method 1300 includes printing the reference electrode (block 1312). In some examples, the rotary screen printer prints the reference electrode onto the base substrate (or into a gap in the base substrate). For example, in device 100 of FIG. 1, the third electrode 124 may be a reference electrode. The third electrode 124 is disposed on the upper surface 112 of the paper substrate 102. The difference between the pair of the first and second electrodes 120, 122 (for example, the working / counter electrode pair) can be measured against the pair of the second and third electrodes i 22, 124 (eg, the reference electrode / counter electrode pair). Similarly, in illustrative device 1100 of FIG. 11, the third electrode 1126 may be a reference electrode. In some examples, the difference between the two pairs of electrodes is indicative of the presence, absence, and / or concentration of the target analyte. In illustrative assembly 500, the first and / or second print station 508, 510 are used to print the reference electrode onto the base substrate 506. In some examples, the working and counter electrode may include a different material than the reference electrode. reference. In such a case, one of the first ^or second printing station 508, 510 may include a first conductive ink (for example, A ^u ) for printing the working electrode and counter electrode, and the other of the first or second printing station Printing 508, 510 may include a second conductive ink (eg, Ag / AgCl) for the reference electrode. The printing of the reference electrode can be controlled by means of, for example, the printer controller (s) 610.

Illustrative method 1300 includes printing a trace and a contact for each of the electrodes (block 1314). In some examples, a trace (eg, a wire ^or cable) is printed on the base substrate ( ^or into a gap in the base substrate) between one of the electrodes and a corresponding contact. The contacts are used to electrically couple the electrodes (which may be covered with hydrophilic ink and a sealing layer) to a reader that can detect electrical signals, including, for example, signals indicative of change / s in volts, amps, and / or or resistance across the electrodes. In some examples, the electrode contacts are arranged near a similar edge of the base substrate, so that the edge can be used with the reader that can easily contact all of the contacts at the same time. For example, device 100 of FIG. 1 includes traces 126, 130, 134 and contacts 128, 132, 136 to the respective electrodes 120, 122, 124. T ^he traces 126, 130, 134 and contacts 128, 132, 136 are disposed on the upper surface 112 of the paper substrate 102. In the illustrated example of FIG. 1, contacts 128, 132, 136 are disposed along the second end 106 of paper substrate 102. As illustrated in FIG. 4, the second end 106 can then be inserted into the reader 400. Similarly, the illustrative device 1100 includes traces 1128, 1132, 1136 and contacts 1130, 1134, 1138 for the respective first, second, and third electrodes 1122, 1124, 1126. . In illustrative assembly 500, the first and / or second print stations 508, 510 can be used to print trace I ^or / ^s and the contact (s) onto the base substrate 506 (or into a gap in the substrate base 506). In some examples, the trace (s) and the contact (s) are printed during the same printing stage as the electrodes. In other examples, they are printed separately. Printing of ^the trace (s) and contact (s) can be controlled by means of, for example, the printer driver (s) 610.

Illustrative method 1300 includes determining whether one ^or more reagents will be included with the device (block 1316). For example, with IA and QC tests, one or more reagents are typically used to react with the sample. If a reagent is to be used, illustrative method 1300 includes the addition of the reagent (block 1318). In some examples, one or more reagents can be printed on the working electrode as reactive ink. Additionally or alternatively, one ^or more reagents may be printed around the electrodes and / or along the area where the hydrophilic ink channel is to be printed. For example, in illustrative device 100 of FIGS. 1 and 2A-2F, reagent 200 is printed on top of first electrode 120. When the sample comes into contact with first electrode 120, the sample interacts with the reagent. In illustrative assembly 500 of FIG. 5, a reagent is printed on the base substrate 506 in the third printing station 512. In some examples, the third printing station 512 is implemented as an inkjet printer. The addition of the reagent (s) can be controlled by, for example, the 61o printer driver (s).

Illustrative method 1300 includes printing a hydrophilic ink or suspension material layer on and around the electrodes (block 1320). The hydrophobic ink layer on top of the electrodes forms a detection zone, where the sample can wet the electrodes and therefore the electrodes can detect the biomolecular interaction that occurs. In some examples, the hydrophilic ink includes particles (eg, microbeads or nanobeads) that cause the sample to pass through the ink material and thus spread across the electrodes. For example, in illustrative device 100 of FIG. 1, the detection zone 116 includes the hydrophobic ink layer 138, which is disposed on and around the electrodes 120, 122, 124 on the upper surface 112 of the paper substrate 102. The layer 138 picks up (for example, by action capillary) a fluid sample through the layer for the sample to contact the reagent and electrodes 120, 122, 124. Similarly, in the illustrative IlOO device, layer 1140 is deposited in gap 1116 over the electrodes 1122, 1124, 1126 to form detection zone 1116. In illustrative assembly 5OO of FIG. 5, the hydrophilic ink layer can be printed on the base substrate 5O6 ( ^or within a gap in the base substrate) at the fourth printing station 514. In the illustrated example, the fourth printing station 514 is implemented as a printer of rotary screen printing. Printing of the hydrophilic ink layer can be controlled, for example, by means of printer controller / s 61O.

The illustrative method 13OO includes printing a channel of hydrophilic ink or suspension material from a first end of the base substrate (or at positions on a roll of substrate that will become one end once the roll is cut in separate devices) to the hydrophilic ink layer (block 1322). The channel provides a pathway for the sample to transfer to the hydrophilic ink layer in the detection zone. In some examples, the channel is formed by a substantially straight path leading from a first end of the base substrate to the layer. Other examples include multiple channels, branched channels, and / or channels with one or more curved portions. A sample deposited on / within the channel will be transported to the layer and, therefore, to the electrodes. For example, device 1OO of FIG. 1 includes channel 14O, which has first end 142 and second end 146 connecting to layer 138. Similarly, in illustrative device 11OO, channel 1142 is deposited in gap 1116 to form channel 1140. In illustrative assembly 5OO of FIG. 5, the hydrophilic ink channel can be printed on the base substrate by means of the fourth printing station 514. In some examples, the ink in the channel and the layer are the same, and the channel and the layer can be printed on the base substrate 5O6 at the same time using rotary screen printing printer. Channel printing can be controlled by, for example, the printer driver (s) 61O.

Illustrative method 13OO includes printing a seal layer over the hydrophilic ink layer and channel (block 1324). The seal layer is a fluid impermeable or hydrophobic layer that reduces evaporation of the sample and / or the risk of contamination. In some examples, the seal layer is printed over the top and sides of the hydrophilic layer and the hydrophilic channel to define a fluid-tight pathway created by the seal layer (e.g., the upper boundary) and the substrate. base (for example, the bottom layer), with the layer and the channel in between. For example, device 1OO of FIG. 1 includes seal layer 148. FIGS. 2F, 3A, and 3B illustrate the arrangement of sealing layer 148 over layer 138 and channel 14O. In illustrative device 11OO, channel 1l42 and detection zone 1118 are formed within gap 1116 in substrate 1102. In that example, seal layer 1150 may be printed over channel 1142 and detection zone 1118 to form the hydrophilic channel between the sealing layer and the surfaces of the gap 1116. In illustrative set 5 ^or 0 of FIG. 5, for example, the sealing layer can be printed on the base substrate 5O6 by means of the fifth printing station 516. In some examples, the fifth printing station 516 is implemented by a rotary screen printer. Printing of the seal layer can be controlled by means of, for example, the printer driver (s) 61O.

Illustrative method 13OO includes separating the base substrate into individual devices (block 1326). In some examples, the method is implemented using an R2R or coil-based manufacturing process. Multiple devices can be printed on the continuous base substrate. The base substrate can be cut ^or diced on the individual devices. In illustrative assembly 5OO of FIG. 5, the optional cutting station 518 is used to cut ^or saw the base substrate 5O6 on the individual devices. The separation ^or dicing can be controlled by means of, for example, the driver (s) 614 of the cutting station.

At the end of Illustrative method 1300 (block 1328), the Individual device can be used to perform, for example, AI ^or CC tests on the base substrate. The resulting device is a relatively inexpensive and disposable diagnostic device that can provide fast and reliable results in a POC setting.

FIG. 14 depicts a representative illustrative flow chart of an alternative illustrative method 1400 for creating a substrate diagnostic device such as, for example, device 700, using, for example, assembly 900 illustrated in FIG. 9. Illustrative method 1400 includes unwinding a first substrate (block 1402). The first substrate acts as a support to build the diagnostic device. In the illustrated example, the first substrate is a hydrophilic sheet of paper (eg, a continuous sheet of paper). The hydrophilic paper sheet absorbs liquids such as, for example, inks. The hydrophilic substrate may correspond to, for example, the paper substrate 702 of the device 70 and / or the base substrate 906 in the illustrative assembly 900. Unwinding of the first substrate can be controlled, for example, by means of roller controller / s 1004.

Illustrative method 1400, which is not in accordance with the present invention, includes printing a hydrophobic barrier on the first substrate to define an outer boundary of a channel and / or detection zone (block 1404). The hydrophobic barrier is printed using a hydrophobic ink. The hydrophobic ink can be printed on the first substrate in a pattern that defines the outer boundary of a channel and / or a detection zone (disclosed herein). The hydrophobic ink absorbs into the material of the first substrate and, as a result, creates a pathway or channel where fluid can be captured by the first substrate. For example, in device 700 of FIG. 7, hydrophobic barrier 738 is a hydrophobic ink line that is disposed on substrate 702. Illustrative barrier 738 defines the outer boundary of channel 742 and detection zone 718, and prevents a fluid sample from being captured in areas of the substrate 702 outside of barrier 738. In illustrative assembly 900, a hydrophobic ink is printed on substrate 906 at first printing station 908. Barrier printing can be controlled by means of, for example, the controller (s). It is 1010 printer.

In illustrative method 1400, a working electrode and a counter electrode on the first substrate (block 1406). The working electrode and counter electrode form an electrical sensor that can be used to determine the presence, absence, and / or concentration of a target analyte in a sample. Different electrical techniques such as amperometric, voltammetric and / or potentiometric techniques can be implemented. The working electrode and counter electrode can be printed on the first substrate, using, for example, conductive inks (eg, metallic). The conductive inks are absorbed into the first substrate and form metal electrodes within the material of the first substrate. For example, on FlG device 700. 7, first electrode 720 and second electrode 722 are printed on paper substrate 702. In the illustrative processing assembly of FIG. 9, the working electrode and / or counter electrode are printed on the substrate 906 in the third printing station 912, which is implemented in an ink jet printer. The inkjet printer can print AuNP ink which is used to form the working electrode and counter electrode. The printing of the electrode can be controlled by means of, for example, the printer controller (s) 1010. In other examples, other types of sensors can be printed on the first base. For example, some devices may include an optical ^or magnetic sensor, and the components thereof may be deposited on the first substrate (or within a gap in the first substrate).

Illustrative method 1400 includes determining if a reference electrode is to be used (block 1408). In some examples, a reference electrode is used, and the difference between the working electrode / counter electrode pair and the reference electrode / counter electrode pair is used to determine the change across the electrodes. The difference can be correlated with the change produced by the sample and the reagent and, therefore, is indicative of the presence, absence and / or concentration of the target analyte. If a reference electrode is to be added, illustrative method 1400 includes printing the reference electrode (block 1410). For example, in device 700 of FIG. 7, the third electrode 724 may be a reference electrode. The third hydrophobic barrier 724 is disposed on the paper substrate 702 and absorbed into the material of the paper substrate 702. In the illustrative assembly 900, the second print station 910 is used to print the reference electrode on the base substrate 906 In the illustrated example of FIG. 9, the second printing station 910 is implemented as a rotary screen printer and includes a conductive ink such as, for example, Ag / AgCl. The printing of the reference electrode can be controlled by means of, for example, the printer controller (s) 1010.

Illustrative method 1400 includes printing a trace and a contact for each of the electrodes on the first substrate (block 1412). In some examples, a trace (eg, a wire ^or cable) is printed on the first substrate between one of the electrodes and a corresponding contact. T ^he contacts are used to electrically couple the electrodes (which may be disposed within the material of the first substrate) to a reader that can detect electrical signals including, for example, signals indicative of change / s in volts, amps and / or resistance through the electrodes. In some examples, the electrode contacts are arranged near a similar edge of the first substrate, so that the edge can be used with the reader that can easily contact all of the contacts at the same time. For example, device 700 of FIG. 7 includes traces 726, 730, 734 and contacts 728, 732, 736 for the respective electrodes 720, 722, 724. Traces 726, 730, 734 and contacts 728, 732, 736 are printed on substrate 702 through conductive inks, and the conductive inks are absorbed into the substrate material 702 to define the traces and contacts. In the illustrative assembly 900, the second and / or third print station 910, 912 can be used to print trace / s. and the contact (s) on the base substrate 906. In some examples, the trace (s) and the contact (s) are printed during the same Printing step as the electrodes. In other examples, they are printed separately. Printing of the trace / s and contact / s can be controlled by means of, for example, the printer driver (s) 1010.

Illustrative method 1400 includes sintering the electrodes (block 1414). In some examples, pressure and / or heat is applied to one or more of the electrodes to fuse the conductive ink atoms and molecules to form a solid material on the first substrate. For example, assembly 900 includes sintering station 914 that is capable of sintering printed electrodes on substrate 906. Sintering station 914 can operate by applying alternating current or direct current to the electrode material. Sintering can be controlled by means of, for example, the conductor (s) 1014 of the sintering station.

Illustrative method 1400 includes determining whether one or more reagents will be included with the device (block 1416). For example, with IA and QC tests, one or more reagents are typically used to react with the sample. If a reagent is to be used, illustrative method 1300 includes the addition of the reagent (block 1418). In some examples, one ^or more reagents can be printed on the working electrode using a reactive ink. Additionally ^or alternatively, one ^or more reagents may be printed around the electrodes and / or along the area where the hydrophilic ink channel is to be printed. In illustrative assembly 900 of FIG. 9, a reagent is printed on the substrate 906 in the fourth printing station 916, which is implemented as an ink jet printer. The addition of the reagent (s) can be controlled by, for example, the printer driver (s) 1010.

Illustrative method 1400 includes printing a hydrophilic mesh layer between the hydrophobic barrier (block 1420). The hydrophilic mesh layer helps capture a fluid sample through the first substrate material. The hydrophilic mesh layer can be printed onto the first substrate using a suspension of hydrophilic material ^or ink that includes particles (eg, microbeads ^or nanobeads) that cause the sample to be captured through the ink material. The hydrophilic mesh layer can define the channel and detection zone of the device. For example, in illustrative device 700 of FIG. 7, the hydrophilic mesh is disposed between the hydrophobic barrier 738 and defines a pathway for a fluid sample to be captured through the channel 742 to the layer 740 of the detection zone. In illustrative assembly 900 of FIG. 9, the hydrophilic mesh layer is printed on the base substrate 906 by means of the mesh laminating unit 918. Mesh printing can be controlled, for example, by means of the 1018 mesh lamination controller (s).

Illustrative method 1400 includes unwinding a second substrate (block 1422). In this example, the second substrate comprises a hydrophobic material, as previously disclosed. In some examples, the second substrate must be attached to one side (eg, the top) of the first substrate and forms a sealing layer or fluid impermeable layer over at least the channel and detection zone of the device. For example, in illustrative device 700 of FIG. 7, the sealing layer 746 is disposed over at least the detection zone 716. In illustrative assembly 900 of FIG. 9, the hydrophobic substrate 922 unwinds from the third roll 920. The unwind of the second substrate can be controlled by means of, for example, the roll driver (s) 1004.

Illustrative method 1400 includes cutting the second substrate and / or creating samples and / or reagent inlet ports on the second substrate (block 1424). In some examples, the second substrate is cut into a shape that substantially matches the shape of the channel and / or the detection zone, so that when the second substrate engages the top of the first substrate, a waterproof seal is created. to the fluids between the second substrate and the hydrophobic barriers on the first substrate. In some examples, a laser is used to cut the second substrate. Additionally or alternatively, the laser can be used to change the hydrophobicity of the second substrate to create sample and / or reagent inlet ports. In illustrative assembly 900 of FIG. 9, the laser 924 is included to cut and / or create entry ports for the sample and / or reagent. The laser can change the hydrophobicity of the hydrophobic substrate 922, so that a sample and / or a reagent can pass through the hydrophobic substrate to reach the channel and / or detection zone on the device. The cutting of the second substrate can be controlled by means of, for example, the controller (s) 1022 of the laser.

Illustrative method 1400 also includes combining the first substrate and the second substrate (block 1426). The second substrate can be arranged on top of the first substrate to seal the top of the first substrate. The second substrate helps prevent evaporation of the sample and / or reduce the risk of contamination. In illustrative assembly 900 of FIG. 9, the lamination unit 926 combines the hydrophobic substrate 922 and the substrate 906. In some examples, the lamination unit 926 uses heat and / or pressure to couple the hydrophobic substrate 922 to the top of the substrate 906. The combination of the primer substrate and the second substrate can be controlled by means of, for example, the controller (s) 1026 of the laminating unit.

Illustrative method 1400 includes separating the combined substrates into individual devices (block 1428). In some examples, the method is implemented using an R2R or coil-based manufacturing process. Multiple devices can be printed on the first and second continuous substrates. The combined substrate can be cut or diced on the individual devices. In illustrative assembly 900 of FIG. 9, the cutting station 928 is used to cut ^or saw the substrate 906 into the individual devices. The separation of the combined substrates in a single device can be controlled by means of, for example, the conductor (s) 1030 from the cutting station.

At the end of illustrative method 1400 (block 1430), individual devices can be used to perform, for example, AI or CC tests on the first substrate. The resulting device is a relatively inexpensive, consumable, and disposable diagnostic device that can provide fast and reliable results in a POC setting. FIG. 15 is a block diagram of an illustrative processor platform 1500 capable of executing the instructions of FIGS. 13 and 14 to implement illustrative assembly 500, illustrative processing system 600, illustrative assembly 900, and / or illustrative processing system 1000 of FIGS. 5, 6, 9, and 10. The processor platform 1500 may be, for example, a server, a personal computer, a mobile device (eg, a cell phone, a smartphone, a tablet such as an iPad ™), a Personal Digital Assistant (PDA), an Internet device, a DVD player, a CD player, a digital video recorder, a Blu-ray player, a game console, a personal video recorder, a decoder ^or any other type of computing device.

The illustrated example processor platform 1500 includes a processor 1512. The illustrated example processor 1512 is hardware. For example, processor 1512 can be implemented by one or more integrated circuits, logic circuits, microprocessors, or controllers from any desired family or manufacturer.

Processor 1512 of the illustrated example includes local memory 1513 (eg, a cache memory). The processor 1512 of the illustrated example is in communication with a main memory that includes a volatile memory 1514 and a non-volatile memory 1516 through a bus 1518. The volatile memory 1514 can be implemented by the synchronous dynamic random access memory (SDRAM, Synchronous Dynamic Random Access Memory), Dynamic Random Access Memory (DRAM), RAMBUS Dynamic Random Access Memory (RDRAM, RAMBUS Dynamic Random Access Memory) and / or any other type of random access memory device . Non-volatile memory 1516 can be implemented by flash memory and / or any other type of memory device desired. Access to main memory 1514, 1516 is controlled by a memory controller.

The processor platform 1500 of the illustrated example also includes an interface circuit 1520. The interface circuit 1520 can be implemented using any type of interface pattern, such as an Ethernet interface, a Universal Serial Bus (USB), and / or a PCI express interface.

In the illustrated example, one ^or more input devices 1522 are connected to interface circuit 1520. EI / I ^os device / s input 1522 allows / na a user to enter data and commands into the processor 1512. EI / device Ios / s input can be implemented by, for example, an audio sensor, a microphone, a camera ( still photo or video), a keyboard, a button, a mouse, a touch screen, a trackpad, a scroll wheel, isopoint and / or a voice recognition system.

One or more output devices 1524 are also connected to the interface circuit 1520 of the illustrated example. T ^he output devices 1524 can be implemented, for example, by display devices (for example, a light emitting diode (LED, Light Emitting Diode), an LED light organic (OLED, Organic Light Emitting Diode), a liquid crystal display, a cathode ray tube (CRT) display, a touch screen, a touch output device, a printer and / or speakers). The interface circuit 1520 of the illustrated example, therefore, typically includes a graphics controller card, a graphics controller chip, ^or a graphics controller processor.

The interface circuit 1520 of the illustrated example also includes a communication device such as a transmitter, receiver, transceiver, modem, and / or network interface card to facilitate data exchange with external machines (eg, devices. of any kind) over a 1526 network (eg, Ethernet connection, Digital Subscriber Line (DSL), telephone line, coaxial cable, cellular phone system, etc.).

The processor platform 1500 of the illustrated example also includes one ^or more mass storage devices 1528 for storing software and / or data. Examples of such 1528 mass storage devices include floppy drives, hard disk drives, compact disk drives, Blu-ray disk drives, RAID systems, and digital versatile disk (DVD) drives.

The encoded instructions 1532 of FIG. 15 can be stored in mass storage device 1528, volatile memory 1514, non-volatile memory 1516, and / or a removable tangible computer-readable storage medium such as a CD or DVD.

From the foregoing, it will be appreciated that the apparatus, systems and I I ^{I I} previously disclosed methods provide disposable diagnostic devices, easily manufacturable and less costly. It is advantageous to use the paper substrate (s) disclosed above, because paper is an easily obtainable, cheap, thin and light material (for example, about 10mg / cm2); it is easy to store and / or transport; has many properties (for for example, the ability to control hydrophobicity and surface characteristics); it is flexible and compatible with many printing methods; it is chemically modifiable to immobilize biomolecules; and it is compatible with colorimetric tests or electrochemical tests using printed electronic circuits. Furthermore, it is advantageous to use the hydrophobic or fluid-impermeable paper substrate (s) disclosed above because the paper can be formed using any substance that does not interact with biological reactions, but is impermeable to aqueous fluids; microfluidic components and sensing components can be deposited into continuous roll-fed processing using printing technologies; In some examples, the process is additive, reducing material consumption and device cost; sealing the microfluidic channel (s) increases the speed at which the fluid flows through the microfluidic channels and, therefore, the device; and the addition of the seal layer forms a defined space volume to aid in fluid measurement.

Although certain illustrative methods, apparatus, systems, and articles of manufacture have been disclosed herein, the scope of the present patent is not limited thereto. On the contrary, the present patent encompasses all the methods, apparatus and systems that are within the scope of the claims of the present patent.

Claims (15)

re iv in d ic a tio ns 1. An apparatus comprising: a hydrophobic substrate (102) having a first end (104) and a second end (106) opposite the first end (104); a detection zone (116) on a first surface (112) of the substrate (102), the detection zone (116) defining an area for detecting an analyte in a sample, the detection zone (116) comprising: a first electrode (120) and a second electrode (122) disposed on the first surface (112) of the substrate (102); and a hydrophilic ink layer (138) disposed on the first and second electrodes (120, 122) and an area between the first and second electrodes (120, 122); and a channel (140) comprising hydrophilic ink disposed on the first surface (112) of the substrate (102), the channel (140) having an inlet section (i 42) adjacent the first end (104) of the substrate (112), a central section (144) and an outlet section (146) in contact with the hydrophilic ink layer (138), which is arranged on the sensor (118), channel (140) that serves to transfer a fluid sample from the inlet section (142) to hydrophilic ink layer (138). The apparatus of claim 1 further comprising a reagent layer (200) disposed on an upper surface of the first electrode (120), between the first electrode (12) and the hydrophilic ink layer (138). The apparatus of claim 1, wherein the first electrode (120) is functionalized with a reagent. 4. The method of claim 1, wherein the hydrophilic ink comprises microbeads or nanobeads and a binder to adhere the microbeads ^or nanobeads to the first surface (112) of the substrate (102). The apparatus of claim 4, wherein the binder comprises at least one of polyvinyl chloride (PVC), polyvinylpyrrolidone (PVP), ^or nanofibrillar cellulose (NFC). 6. A system for detecting an analyte in a sample, the system comprising: the apparatus of claim 1, further comprising: a first contact (128) disposed on the first surface (112) of the substrate (102), the first contact (128) being electrically coupled to the first electrode (120); and a second contact (132) disposed on the first surface (112) of the substrate (102), the second contact (132) being electrically coupled to the second electrode (122); and a reader (400) comprising: a slot (402) for receiving at least a portion of the substrate (102) including the first contact (128) and the second contact (132); electrical connectors for receiving signals from the first contact (128) and the second contact (132); and a processor to determine one or more of amperometric, voltammetric, or potentiometric measurements to detect a biomolecular interaction occurring in the detection zone (116) through the first and second electrodes (120, 122). The system of claim 6, wherein the detection zone (116) further comprises a third electrode (124) and the processor must detect the biomolecular interaction by comparing (1) a difference in the first and second electrodes (120, 122) and (2) a difference in the second and third electrodes (122, 124). The system of claim 6, wherein the detection zone (116) comprises a plurality of additional electrodes, and the processor must determine a plurality of measurements using the plurality of additional electrodes. The apparatus of claim 1, wherein the substrate (102) comprises hydrophobic paper. The apparatus of claim 1, wherein the substrate (102) is flexible and printable. 11. A method of manufacturing a diagnostic device on substrate, the method comprising: depositing a sensor (118), including a first electrode (120) and a second electrode (122), on an upper surface (112) of a hydrophobic substrate (102), the hydrophobic substrate (102) having a first end (104) and a second end (106) opposite the first end (104); depositing a hydrophilic ink layer (138) on the sensor (118), the sensor (118) and the hydrophilic ink layer (138) forming a detection zone (116) for detecting an analyte in a sample; and depositing a path (140) of hydrophilic ink on the upper surface (112) of the substrate (102), the path (14o) leading from an area adjacent to the first end (104) of the substrate (102) to the layer (138) of hydrophilic ink. The method of claim 11, wherein deposition of the sensor comprises printing the first electrode (120) and the second electrode (122) on the upper surface (112) of the substrate (1o2). The method of claim 12 further comprising depositing a reagent layer (200) on the first electrode (120), wherein the reagent layer (200) is disposed between the first electrode (120) and the layer (i 38 ) of hydrophilic ink. The method of claim 12, wherein the first electrode (120) is functionalized with a reagent. The method of claim 11, wherein the deposition of the hydrophilic ink layer (138) and the deposition of the hydrophilic ink path (140) comprises printing the hydrophilic ink using at least one of a flexographic printer, a printer screen printing, a stencil printer or an inkjet printer.